An English Medium Co. Ed.

Senior

Secondary School

Investigatory
Project
On

SUBM ITTE  B!"

SUBMITTE TO"
Su#hag Singh Mr.
Sandee$ %ulshesthra
&II Sci. B
'(.O. Biology)

A*no+ledgement
I am over+helmed in all hum#leness and grate,ulness to ac*no+ledge
my de$th to all those +ho have hel$ed me to $ut these ideas- +ell
a#ove the level o, sim$licity and into something concrete.
I +ould li*e to e$ress my s$ecial than*s o, gratitude to my #iology
teacher- Mr. Sandeep Kulshesthra as +ell as our Princi$al Mrs. Nidhi
Bhatia +ho gave me the golden o$$ortunity to do this +onder,ul
$roject on the to$ic /A$$lications o, Biotechnology0- +hich also hel$ed
me in doing a lot o, research and I came to *no+ a#out so many ne+
things. I am really than*,ul to them.
Any attem$t at any level can1t #e satis,actorily com$leted +ithout the
su$$ort and guidance o, my Parents and Friends +ho hel$ed me a
lot in gathering di2erent in,ormation- collecting data and guiding me
,rom time to time in ma*ing this $roject- des$ite o, their #usy
schedules- they gave me di2erent ideas in ma*ing this $roject
uni3ue. I am than*,ul to them too.
I am ma*ing this $roject not only ,or mar*s #ut to also increase my
*no+ledge...
 Than*ing you
Subhag Singh
XII Sci. B
 Certi4cate
 This is to certi,y that SUBHAG SINGH o,
class XII SCI.B o, GAN !""P SHIKSHA
BHA#A$I
has success,ully com$leted the investigatory
$roject on the to$ic %APP&ICA$I'NS 'F
BI'$"CHN'&'G( under the guidance
o, M#.
SAN!""P KU&SH"S$H#A  '(.O..
Biology) during the session 567897: in the
$artial ,ul4lment o, Biology Practical
Eamination conducted #y C"N$#A& B'A#!
'F S"C'N!A# "!UCA$I'N )AISSC"*.
 ;;;;;;;;;;;;;;;;;;;
 ;;;;;;;;;;;;;;;;;;; 
Mr. Sandee$ %ulshesthra Eternal
Eaminer
'(.O. Biology) 'C.B.S.E)

Introduction
+hat is Bi,techn,l,g-

Bi,techn,l,g- is the use o, living systems and organisms to develo$

or ma*e $roducts- or <any technological a$$lication that uses
#iological systems- living organisms or derivatives thereo,- to ma*e
or modi,y
$roducts or $ rocesses , s $eci4c u se.
or

 At its sim$lest- #iotechnology

is technology #ased on #iology 9
#iotechnology harnesses cellular
and #io molecular
$rocesses to develo$
technologies and
$roducts that hel$
im$rove our lives and the
health o, our $lanet.
=e have used the #iological
$rocesses o, microorganisms ,or
more than :-666 years to ma*e
use,ul ,ood $roducts- such as #read and cheese- and to $reserve dairy
$roducts.

Modern #iotechnology $rovides #rea*through $roducts and

technologies to com#at de#ilitating and rare diseases- reduce our
environmental ,oot$rint- ,eed the hungry- useless and cleaner energy-
and have sa,er- cleaner and more e>cient industrial manu,acturing
$rocesses.
 Biotech is hel$ing to heal the +orld #y harnessing nature?s o+n tool#o
and using our o+n genetic ma*eu$ to heal and guide lines o, research
#y"
• @educing rates o, in,ectious disease
• Saving millions o, children?s lives
• Changing the odds o, serious- li,e9threatening conditions a2ecting
millions around the +orld
•  Tailoring treatments to individuals to minimie health ris*s
and side e2ects
• Creating more $recise tools ,or disease detection
• Com#ating serious illnesses and everyday threats con,ronting the
develo$ing +orld.

(istory
 Throughout the history o, agriculture- ,armers have inadvertently
altered the genetics o, their cro$s through introducing them to ne+
environments and #reeding them +ith other $lants 9 one o, the 4rst
,orms o, #iotechnology.
 These $rocesses also +ere included in early /er0entati,n ,/ beer.
In #re+ing- malted grains 'containing enymes) convert starch ,rom
grains into sugar and then adding
s$eci4c yeasts to $roduce #eer. In
this $rocess- car#ohydrates in the
grains +ere #ro*en do+n into
alcohols such as ethanol. ater
other cultures $roduced the
$rocess o, lactic acid
,ermentation +hich allo+ed the
,ermentation and $reservation
o, other ,orms o, ,ood- such as
s,- sauce. ermentation +as
also
used in this time $eriod to $roduce lea1ened bread. Although the
$rocess o, ,ermentation +as not ,ully understood until ouis Pasteur?s
+or* in 7D8- it is still the 4rst use o, #iotechnology to convert a ,ood
source into another ,orm.

or thousands o, years- humans have used selective #reeding

to im$rove $roduction o, cro$s and livestoc* to use them ,or ,ood. In
selective #reeding- organisms +ith desira#le characteristics are mated
to $roduce o2s$ring +ith the same characteristics. or eam$le- this
techni3ue +as used +ith corn to $roduce the largest and s+eetest
cro$s.

Biotechnology has also led to the develo$ment o, anti#iotics. In 7F5D-

Aleander leming discovered the mould Penicilliu0. (is +or* led to
the $uri4cation o, the anti#iotic com$ound ,ormed #y the mould #y
(o+ard lorey- Ernst Boris Chain and Gorman (eatley 9 to ,orm +hat
+e today *no+ as $enicillin. In 7FH6- $enicillin #ecame availa#le ,or
medicinal use to treat #acterial in,ections in humans.

 The 4eld o, modern #iotechnology is generally thought o, as having

#een #orn in 7F7 +hen Paul Berg?s e$eriments in gene splicing had
early success. (er#ert =. Boyer and Stanley G. Cohen
signi4cantly advanced the ne+ technology in 7F5 #y trans,erring
genetic material into a #acterium- such that the im$orted material +ould
#e re$roduced.
 Biotechnology in
Agriculture
Geneticall- M,di2ed Cr,ps

Geneticall- 0,di2ed cr,ps or

/M cro$s0 or /#iotech cro$s0 are
$lants used in agriculture- the GA o,
+hich has #een modi4ed +ith genetic
engineering techni3ues. In most cases
the aim is to introduce a ne+ trait to
the
$lant +hich does not occur naturally
in the s$ecies. Eam$les in ,ood
cro$s
include resistance to certain $ests- diseases- stress,ul environmental
conditions- resistance to chemical treatments- reduction o, s$oilage- or
im$roving the nutrient $ro4le o, the cro$. Eam$les in non9,ood cro$s
include $roduction o, $harmaceutical agents- #io ,uels- and
other industrially use,ul goods- as +ell as ,or #ioremediation.

Plants and cro$s +ith M traits have #een tested more than any
other cro$sJ+ith no credi#le evidence o, harm to humans or animals.
In ,act- seeds +ith M traits have #een tested more than any other
cro$s in the history o, agriculture K +ith no credi#le evidence o,
harm to humans or animals.

overnmental regulatory agencies- scienti4c organiations and leading

health associations +orld+ide agree that ,ood gro+n ,rom GM cr,ps is
sa/e t, eat. The =orld (ealth Organiation- the American
Medical Association- the U.S. Gational Academy o, Sciences- the
British @oyal Society- among others that have eamined the evidence-
all come to the same conclusion" consuming ,oods containing
ingredients derived ,rom
M cro$s is sa,e to eat and no ris*ier than consuming the same ,oods
containing ingredients ,rom cro$ $lants modi4ed #y conventional $lant
im$rovement techni3ues. enetic modi4cations have"

7. Made cro$s more tolerant to a#iotic stresses 'cold- drought- salt-

heat).
5. @educed reliance on chemical $esticides '$est resistant cro$s).
 L. (el$ed to reduce $ost harvest losses  enhanced the nutritional
value o, the ,ood.

#NA Inter/erence )#NAi*

@GA inter,erence '@GAi) is a method o, #loc*ing gene ,unction
#y inserting short se3uences o, ri#onucleic acid '@GA) that match $art
o, the target gene1s se3uence- thus no $roteins are $roduced. @GAi has
the
$otential to #ecome a $o+er,ul thera$eutic a$$roach to+ard targeted
and $ersonalied medicine. @GAi has $rovided a +ay to control $ests
and diseases- introduce novel $lant traits and increase cro$ yield. Using
@GAi- scientists have develo$ed novel cro$s such as
nicotine9,ree to#acco- non9allergenic $eanuts- deca2einated co2ee-
and nutrient
,orti4ed maie among many others.

Mechanism o, @GA inter,erences as understood is that it comes into

$lay
+hen a dou#le stranded @GA is introduced either naturally or
arti4cially in a cell. An endo ri#onuclease enyme cleaves the long
ds@GA into small $ieces o, @GA. The small $ieces could #e mi
@GA or si @GA de$ending u$on the origin o, long ds@GA i.e.
endogenous or eogenous
res$ectively. A dou#le
stranded @GA may #e
generated #y either @GA
de$endent @GA $olymerase
or #idirectional transcri$tion
o, trans$osa#le elements or
$hysically introduced.

 There are several

o$$ortunities ,or the a$
$lications o, @GAi in cro$
science ,or its im$rovement
such as stress tolerance and
enhanced nutritional level.This *noc*do+n technology may #e use,ul in
inducing early No+ering- delayed ri$ening- delayed senescence-
#rea*ing dormancy- stress9,ree $lants- overcoming sel,9sterility- etc .
@GA inter,erence '@GAi) has recently #een demonstrated in $lant
$arasitic nematodes. It is a $otentially $o+er,ul investigative tool ,or the
genome9+ide identi4cation o, gene ,unction that should hel$ im$rove
our understanding o, $lant $arasitic nematodes. @GAi should hel$
identi,y gene and- hence- $rotein targets ,or nematode control
strategies. Pros$ects ,or novel resistance de$end on the $lant
generating an e2ective ,orm o, dou#le9stranded @GA in the a#sence
o, an endogenous target gene +ithout detriment to itsel,. These @GA
molecules must then #ecome availa#le to the nematode and #e ca$a#le
o, ingestion via its ,eeding tu#e. I, these re3uirements can #e met- cro$
resistance could #e achieved #y a $lant delivering a ds@GA that targets
a nematode gene and induces a lethal or highly damaging @GAi e2ect
on the $arasite.

Bt t,3in
A $rotein that is toic to che+ing insects and is $roduced #y the soil
#acterium Bacillus thuringiensis and has long #een used as a #iological
pesticide. By means o, genetic engineering- the genes ,or Bt toin can
#e isolated ,rom Bacillus thuringiensis and trans,erred to $lants.

Bacillus thuringiensis )Bt* is a #acteria that $roduces $roteins +hich are

toic to insects. But etreme toicity comes at no sur$rise. It1s in the
same ,amily o, #acteria as B. anthracis- +hich causes anthra- and B.
cereus- +hich causes ,ood $oisoning.

 The Bt   toin dissolve in the high $( insect gut and #ecome active. The
toins then attac* the gut cells o, the insect- $unching holes in the
lining. The Bt   s$ores s$ills out o, the gut and germinate in the insect
causing death +ithin a cou$le days.

Even though the toin does not *ill the insect immediately- treated $lant
$arts +ill not #e damaged #ecause the insect sto$s ,eeding +ithin
hours. Bt  s$ores do not s$read to other insects or cause disease
out#rea*s on their o+n.
7. Insect eats Bt   crystals and s$ores.

5. The toin #inds to s$eci4c

rece$tors in the gut and the insects
sto$s eating.

L. The crystals cause the gut +all to

#rea* do+n- allo+ing s$ores and
normal gut #acteria to enter the #ody.

H. The insect dies as s$ores and

gut #acteria $roli,erate in the #ody.

Bt   action is very s$eci4c. i2erent strains o, Bt   are s$eci4c to di2erent
rece$tors in insect gut +all. Bt  toicity de$ends on recogniing
rece$tors- damage to the gut #y the toin occurs u$on #inding to a
rece$tor. Each insect s$ecies $ossesses di2erent ty$es o, rece$tors that
+ill match only certain toin $roteins- li*e a loc* to a *ey.

It is #ecause o, this that ,armers have to #e care,ul to match the target

$est s$ecies +ith a $articular Bt   toin $rotein +hich is s$eci4c ,or that
insect. This also hel$s the #eni4cal insects #ecause they +ill usually not
#e harmed #y that $articular strain o, Bt  .

Bt C,tt,n
Bt c,tt,n is a genetically modi4ed organism 'MO) cotton variety-
+hich $roduces an insecticide to #oll+orm. Strains o, the #acterium
Bacillus thuringiensis $roduce over 566 di2erent Bt toins-
each harm,ul to di2erent insects. Most nota#ly-
Bt toins are insecticidal to the larvae o, moths
and #utterNies- #eetles- cotton #oll+orms and
ghtu Nies #ut are harmless to other ,orms o, li,e.
 The gene coding ,or Bt toin has #een inserted
into cotton as a transgene- causing it to $roduce
this natural insecticide in its tissues. In many
regions- the main $ests in commercial cotton
are le$ido$teran larvae- +hich are *illed #y the
Bt $rotein in thegenetically modi4ed cotton they
eat. This eliminates the need to use large
amounts o, #road9s$ectrum insecticides to *ill le$ido$teran $ests.
This s$ares natural insect $redators in the ,arm ecology and ,urther
contri#utes to non insecticide $est management.

Bt cotton is ine2ective against many cotton $ests such as $lant

#ugs- stin* #ugs- and a$hids de$ending on circumstances it may #e
desira#le to use insecticides in $revention. A 566: study
done #y Cornell researchers- the Center ,or
Chinese Agricultural Policy and the Chinese
Academy o, Science on Bt cotton ,arming in
China ,ound that a,ter seven years these
secondary $ests that +ere normally
controlled #y $esticide had increased-
necessitating the use o, $esticides at similar
levels to non9Bt cotton and causing less
$ro4t ,or ,armers #ecause o, the
etra e$ense o, M seeds.

Mechanism:

Bt cotton +as created through the addition o, genes encoding toin

crystals in the Cry grou$ o, endotoin. =hen insects attac* and eat the
cotton $lant the Cry toins are dissolved due to the high $( level o, the
insects stomach. The dissolved and activated Cry molecules #ond to
cadherin9li*e $roteins on cells com$rising the #rush #order
molecules. The e$ithelium o, the #rush #order mem#ranes se$arates
the #ody cavity ,rom the gut +hilst allo+ing access ,or nutrients. The
Cry toin molecules attach themselves to s$eci4c locations on the
cadherin9li*e $roteins $resent on the e$ithelial cells o, the midge and
ion channels are ,ormed +hich allo+ the No+ o, $otassium. @egulation
o, $otassium concentration is essential and- i, le,t unchec*ed-
causes death o, cells. ue to the ,ormation o, Cry ion channels
su>cient regulation o, $otassium ions is lost and results in the death
o, e$ithelial cells. The death o, such cells creates ga$s in the #rush
#order mem#rane.

 Advantages:
Bt cotton has several advantages over non Bt cotton. The im$ortant
advantages o, Bt cotton are #rieNy "

• Increases yield o, cotton due to e2ective control o, three ty$es

o, #oll+orms- vi. American- S$otted and Pin* #oll+orms.

• Insects #elonged to e$ido$tera 'Boll+orms) are sensitive

to crystalline endotoic $rotein $roduced #y Bt gene +hich in
turn
$rotects cotton ,rom #oll+orms.

• @eduction in $esticide use in the cultivation o, Bt cotton in +hich

#oll+orms are major $ests.

• @eduction in the cost o, cultivation and lo+er ,arming ris*s.

• @eduction in environmental $ollution #y the use o, insecticides

rarely.

• Bt cotton ehi#it genetic resistance or in#uilt resistance +hich is a

$ermanent ty$e o, resistance and not a2ected #y environmental
,actors. Thus $rotects cro$ ,rom #oll+orms.

• Bt cotton is eco,riendly and does not have adverse e2ect on

$arasites- $redators- #ene4cial insecticides and organisms $resent in
soil.
• It $romotes
multi$lication o,
$arasites and $redators
+hich hel$ in
controlling the
#oll+orms #y ,eeding
on larvae and eggs
o, #oll+orm.

• Go health haards
due to rare use o,
insecticides.

• Bt cotton are early in maturing as com$ared to non Bt cotton.

Disadvantages:
Bt cotton has some limitations

• (igh cost o, Bt cotton seeds as com$ared to non Bt cotton seeds.

• E2ectiveness u$ to 756 days- a,ter that the toin

$roducing e>ciency o, the Bt gene drastically reduces.

• Ine2ective against suc*ing $ests li*e jassids- a$hids- +hiteNy etc.

Bt cotton in India:

Bt cotton is su$$lied in India?s Maharashtra state #y the

agri9 #iotechnology com$any- Mahyco- as the distri#utor.

 The use o, Bt cotton in India has gro+n e$onentially since

its introduction. @ecently India has #ecome the num#er one glo#al
e$orter o, cotton and the second largest cotton $roducer in the +orld.
India has #red Bt9cotton varieties such as Bikaneri Nerma and hy#rids
such as G((9HH- setting u$ India to #ene4t no+ and +ell into the ,uture.

India1s success has #een su#ject to scrutiny. Monsanto?s seeds are

e$ensive and lose vigour a,ter one generation- $rom$ting the Indian
Council o, Agricultural @esearch to develo$ a chea$er Bt cotton
variety
+ith seeds that could #e reused. The cotton incor$orated the cry7Ac
gene ,rom the soil #acterium Bacillus thuringiensis 'Bt)- ma*ing
the cotton toic to #oll+orms. In $arts o, India cases o, ac3uired
resistance against Bt cotton have occurred.

 The state o, Maharashtra #anned the sale and distri#ution o, Bt cotton

in 5675- to $romote local Indian seeds- +hich demand less +ater-
,ertiliers and $esticide in$ut- #ut li,ted the #an in 567L.

India a$$roved Bt cotton in 5665 no+ it accounts ,or F5 o, all

Indian cotton. Average nation+ide cotton yields +ent ,rom L65 *gQha
in the 5665QL season to a $rojected HD7 *gQha in 5677Q75 J u$ 8F.L
overall.
 This chart sho+s the trends in yields- +hich too* o2 a,ter Bt +as
introduced in 5665. The gra$hs also sho+ that J and here comes
ugly
,actJ in the last H years- as Bt has risen ,rom : to F5 o, India1s
cotton- yields have dro$$ed steadily.

Biotechnology
in Medicine
Geneticall- "ngineered Insulin )Hu0ulin*
Insulin is a $e$tide hormone $roduced
#y #eta cells in the $ancreas o, various
organisms including human #eings. It
regulates
the 0etab,lis0 ,/ carb,h-drates an
d ,ats #y $romoting the a#sor$tion
o, glucose ,rom the #lood to
s*eletal muscles and ,at tissue and
#y causing
,at to #e stored rather than used ,or energy. Insulin also inhi#its the
$roduction o, glucose #y the liver.

Ece$t in the $resence o, the meta#olic disorder

dia#etes mellitus and meta#olic syndrome- insulin is $rovided +ithin
the #ody in a constant $ro$ortion to remove ecess glucose ,rom the
#lood- +hich other+ise +ould #e toic. =hen #lood glucose levels ,all
#elo+ a certain level- the #ody #egins to use stored glucose as an
energy source through glycogenolysis- +hich #rea*s do+n the
glycogen stored in the liver and muscles into glucose- +hich can then
#e utilied as an energy source. As a central meta#olic control
mechanism- its status is also used as a control signal to other #ody
systems 'such as amino acid u$ta*e #y #ody cells). In addition- it has
several other ana#olic e2ects throughout the #ody. +hen
c,ntr,l ,/ insulin le1els /ails4 diabetes 0ellitus can result.

Structure:

Insulin is com$osed o, t+o

di2erent ty$es o, $e$tide
chains. Chain A has 57 amino
acids and Chain B has L6 amino
acids. Both chains contain al$ha
helices #ut no #eta strands.
 There are L conserved disul2de
bridges +hich hel$ *ee$ the
t+o chains together. Insulin
can also ,orm dimers in
solution due
to the hydrogen #onding #et+een the B chains. The dimers can ,urther
interact to ,orm heamers due to interaction #et+een hydro$ho#ic
sur,aces. This scene highlights the hydro$ho#ic and $olar $arts o, an
insulin monomer at a $( o, .

A num#er o, insulin variants have #een made to ,avor either the

monomeric or heameric ,orm. eletion o, the 4ve C terminal residues
o, the B chain creates a monomer only ,orm. This $ortion o, the B
chain is involved in hydrogen #onds #et+een the B chain o, one
monomer and the A and B chain o, another monomer .

Need of Genetically Engineered Insulin:

 The original ,orm o, the +onder cure ,or dia#etes- these +ere once the
only ty$e o, insulin availa#le- #ut are no+ rarely used. Ani0al insulin
+as originally made
,rom ground9u$
animal $ancreas
tissue- and then later
+as etracted ,rom
healthy animals
'slaughtered $igs 
co+s). The
meta#olism o, co+s and $igs +as close enough to human
meta#olism that their animal insulin also +or*ed +ell in human
#odies. Bee, insulin has L di2erences ,rom human $or* insulin has 7
di2erence ,rom human. The use o, a miture o, #ee, and $or* insulin
+as also $ossi#le.
 It has #een sho+n that human insulin is less immunogenic than animal
insulin. Porcine insulin is most similar to human insulin. The $rimary
amino acid se3uences o, #ovine and $orcine insulin di2er ,rom that
o, human insulin #y three and one amino acid- res$ectively. This greater
dissimilarity #et+een human and #ovine insulin has #een $ostulated to
#e the e$lanation ,or the greater antigenicity o, #ovine insulin as
com$ared +ith $orcine insulin

One o, the pr,ble0s 5ith ani0al insulin 5as antib,d- issues. The
#ody identi4es them and tries to reject them. Por* insulin di2ers
#y 7 amino acid and #ee, insulin #y L amino acids- so the #ody?s
immune system can sometimes recognie them as ,oreign.
Immunological com$lications o, insulin thera$y have #een evident
since animal insulin #ecame availa#le ,or the treatment o, dia#etes
mellitus in 7F55. In insulin9allergic $atients treated +ith
conventional insulin $re$arations-
the insulin9s$eci4c IgE values are o,ten 769 to 569,old higher than in
$atients +ithout allergy. It has #een sho+n that human insulin is less
immunogenic than animal insulin. Porcine
insulin is most similar to human insulin.
Cross9 reactivity #et+een human insulin and
insulin o, animal origin has #een re$orted. A
major
$ro#lem is the cross9reactivity that occurs
#et+een anti9insulin anti#odies and the
various animal and human insulin $re$arations
in $atients $resenting +ith allergy to animal
insulin.

 The usage o, ani0al insulin has s, greatl- declined in 0,dern

ti0es that they have largely #een +ithdra+n ,rom the mar*et.
Ge+ly diagnosed dia#etics are ty$ically given synthesied or
Geneticall- "ngineered hu0an insulin.

What is !roinsulin"#

Pr,insulin is the $rohormone $recursor to insulin made in the

#eta cells o, the islets o, angerhans- s$ecialied regions o, the
$ancreas.
Proinsulin is synthesied on
mem#rane associated
ri#osomes ,ound on the rough
endo$lasmic reticulum- +here it
is ,olded and its disul4de #onds
are oidied. It is then
trans$orted to the olgi a$
$aratus +here it is $ac*aged
into secretory vesicles- and
+here it is $rocessed #y a series
o, $roteases to ,orm
mature insulin. Mature insulin has L8 ,e+er amino acids H are
removed altogether- and the remaining L7 ,orm the C9$e$tide. The
C9$e$tide is a#stracted ,rom the center o, the $roinsulin se3uence the
t+o other ends 'the B chain and A chain) remain connected #y disul4de
#onds.

 =hen insulin +as originally $uri4ed ,rom #ovine or $orcine $ancreata-

all the $roinsulin +as not ,ully removed. RLRH =hen some $eo$le used
these insulins- the $roinsulin may have caused the #ody to react +ith a
rash- to resist the insulin- or even to ma*e dents or lum$s in the s*in at
the $lace +here the insulin +as injected. This can #e descri#ed as
an iatrogenic injury due to slight di2erences #et+een the $roinsulin
o, di2erent s$ecies. Since the late 7F6s- +hen
highly
$uri4ed $orcine insulin +as introduced- and the level o, insulin $urity
reached FF- this ceased to #e a signi4cant clinical issue. $he
0ain challenge /,r pr,ducti,n ,/ insulin using r!NA techni6ues
5as getting insulin asse0bled int, 0ature /,r0.

$umulin:

Hu0ulin +as the 4rst medication $roduced using modern genetic

engineering techni3ues in +hich actual human GA is inserted into
a host cell 'E. coli in this case). Biosynthetic <human< insulin is
no+ manu,actured ,or +ides$read clinical use using genetic
engineering techni3ues using recom#inant GA technology- +hich
the manu,acturers claim reduces the $resence o, many im$urities-
although there is no clinical evidence to su#stantiate this claim .
"li &ill- 0ar7eted the 2rst arti2cial insulin4 Hu0ulin4 in 89:;.

(umulin $roduction method is as ,ollo+s"

7. GA coding ,or A and B $oly$e$tide chains o, insulin are

chemically synthesised a in the la#. Sity three nucleotides are
se3uenced to $roduce A chain o, insulin and ninety nucleotide long
GA designed to $roduce B chain o, insulin- $lus terminator
codon is added at the end o, each chain se3uence. Anti9codon
,or methionine is added at the #eginning o, the se3uence to
distinguish humulin ,rom the other #acterial $roteins.

5. Chemically synthesied A and B chain GA se3uence are inserted

into one o, the mar*er gene +hich are $resent in the $lasmid
vector. enes are inserted into the $lasmid +ith the hel$
o, enymes *no+n as endonuclease and ligase.

L. The vector $lasmids +ith the insulin gene are then introduced into
the E. coli #acterial cell. These cells are then allo+ed to re$licate
#y mitosis- along +ith the #acterial cell recom#inant $lasmid also
gets re$licated $roducing the human insulin.

H. A and B $oly$e$tide chains o, insulin are then etracted and

$uri4ed ,rom the ,omenters in the la#. (igh9Per,ormance i3uid
 Chromatogra$hy '(PC) is used to get 766 $ure humulin ,rom
the miture o, $roteins.

8. The A and B $oly$e$tide chains o, insulin are mied together and

connected +ith each other #y disul$hide #ond- ,orming the
(umulin or synthetic human insulin.

 Advantages % Disadvantages of $umulin:

(umulin is the one and only human $rotein $roduced in the #acteria
+ith identical chemical structure to that o, the natural human insulin.
Administration o, humulin reduces the $ossi#ility o, anti#ody
$roduction and inNammatory res$onse
in dia#etic $atients. Major
di>culty is the etraction
o, humulin ,rom a miture
o, host $roteins $resent in
the
,ermentation #roth.

Go+ days to overcome this

etraction $ro#lem synthetic
human insulin are $roduced
in the yeast cell instead o, E. coli using the same $rocedure. As yeast is
Eu*aryotes they secrete the +hole humulin molecule +ith $er,ect three
dimensional structures- reducing the need ,or com$le and
time consuming $uri4cation methods.

Go+ most o, the dia#etic $atients are treated +ith synthetic human
insulin. Small grou$ o, $atients claim that e$isodes o, hy$erglycaemic
com$lications have #een increased a,ter shi,ting ,rom animal origin
insulin to humulin. Go study till date sho+s the di2erence #et+een the
,re3uency o, hy$erglycaemic com$lications in $atient using humulin
'synthetic human insulin) and animal origin insulin.
Gene $herap-
Gene therap- is the thera$eutic delivery o, nucleic acid $olymers into a
$atient?s cells as a drug to treat disease. ene thera$y is an
e$erimental techni3ue that uses genes to treat or $revent disease. In
the ,uture- this techni3ue
may allo+ doctors to treat a
disorder #y inserting a gene
into a $atient1s cells instead
o, using drugs or surgery.
@esearchers are testing
several a$$roaches to gene
thera$y- including"

• @e$lacing a mutated
gene that causes
disease +ith a healthy
co$y o, the gene.

• Inactivating- or /*noc*ing out-0 a mutated gene that is

,unctioning im$ro$erly.

• Introducing a ne+ gene into the #ody to hel$ 4ght a disease.

Although gene thera$y is a $romising treatment o$tion ,or a num#er

o, diseases 'including inherited disorders- some ty$es o, cancer- and
certain viral in,ections)- the techni3ue remains ris*y and is still under
study to ma*e sure that it +ill #e sa,e and e2ective. ene thera$y is
currently only #eing tested ,or the treatment o, diseases that have no
other cures. It should #e noted that not all medical $rocedures that
introduce alterations to a $atient?s genetic ma*eu$ can #e considered
gene thera$y. Bone mar o+ trans$lantation- and organ trans$lants in
general have #een ,ound to introduce ,oreign GA into $atients. ene
thera$y is de4ned #y the $recision o, the $rocedure and the intention
o, direct thera$eutic e2ects.

ene thera$y +as conce$tualied in 7F5- #y authors +ho urged

caution #e,ore commencing human gene thera$y studies.

 The 4rst attem$t- al#eit an unsuccess,ul one- at gene thera$y 'as +ell
as the 4rst case o, medical trans,er o, ,oreign genes into humans not
counting organ trans$lantation) +as $er,ormed #y Martin Cline on 76
 uly 7FD6. Cline claimed that one o, the genes in his $atients +as active
si months later- though he never $u#lished this data or had
it
veri4ed and even i, he is correct- it?s unli*ely it $roduced any signi4cant
#ene4cial e2ects treating #eta9thalassemia.

 The 4rst germ line gene thera$y consisted o, $roducing a genetically

engineered em#ryo in Octo#er 7FF:. The #a#y +as #orn on uly 57-
7FF and +as $roduced #y ta*ing a donor?s egg +ith healthy
mitochondria- removing its nuclear GA and 4lling it +ith the nuclear
GA o, the #iological mother 9 a $rocedure *no+n as cyto$lasmic
trans,er.

 This $rocedure +as re,erred to sensationally and some+hat

inaccurately in the media as a <three $arent #a#y<- though mtGA is
not the $rimary human genome and has little e2ect on an
organism?s individual characteristics #eyond $o+ering their cells.

ene thera$y is a +ay to 4 a genetic $ro#lem at its source. The

$olymers are either e$ressed as $roteins- inter,ere +ith
$rotein e$ression- or $ossi#ly correct genetic mutations.

 The most common ,orm uses GA that encodes a

,unctional- thera$eutic gene to re$lace a mutated gene. The
$olymer molecule is
$ac*aged +ithin a <vector<- +hich carries the molecule inside cells.

 The 4rst commercial gene thera$y- endicine- +as a$$roved in China

in 566L ,or the treatment o, certain cancers. In 5677 Geovasculgen +as
registered in @ussia as the 4rst9in9class gene9thera$y drug ,or
treatment o, $eri$heral artery disease- including critical lim#
ischemia. In 5675 ly#era- a treatment ,or a rare inherited disorder-
#ecame the 4rst treatment to #e a$$roved ,or clinical use in either
Euro$e or the United States a,ter its endorsement #y the Euro$ean
Commission.

A!A de2cienc- is one ,orm o, SCI 'severe

com#ined immunode4ciency)- a disorder that a2ects the immune
system. AA de4ciency is very rare- #ut very dangerous- #ecause a
mal,unctioning immune system leaves the #ody o$en to in,ection
,rom #acteria and viruses.
 The disease is caused #y a
mutation in a gene on
chromosome 56. A!A
de2cienc- is inherited in
an aut,s,0al
recessi1e 0anner. The
gene codes ,or the
enyme adenosine
deaminase 'AA).
=ithout this enyme- the
#ody is una#le to #rea*
do+n a toic su#stance called
deoyadenosine. The toin
#uilds u$ and destroys
in,ection94ghting
immune cells called T and B
lym$hocytes. Because AA
de4ciency a2ects the
immune system- $eo$le +ho have the disorder are more susce$ti#le to
all *inds o, in,ections- $articularly those o, the s*in- res$iratory system-
and gastrointestinal tract. They may also #e shorter than normal. Sadly-
most #a#ies +ho are #orn +ith the disorder die +ithin a ,e+ months.

$reat0ents ,/ A!A !e2cienc- includes"

• #one marro+ trans$lant

gene therap-

• AA enyme in PE vehicle

'n Septe0ber 8<4 899=4 the 2rst gene therap-  to com#at this
disease +as $er,ormed #y r. =illiam rench Anderson on a /,ur>-ear>
,ld girl4 Ashanti !eSil1a- at the Gational Institutes o,
(ealth- Bethesda- Maryland- U.S.A.

Conclusion
Biotechnology is the ne+ +onder o, science. It is truly
multidisci$linary in nature and it encom$asses several disci$lines o,
#asic sciences and engineering. The Science disci$lines ,rom +hich
#iotechnology dra+s heavily are micro#iology- chemistry-
#iochemistry- genetics- molecular #iology- immunology- cell and
tissue culture and $hysiology. On the engineering side it leans
heavily on $rocess chemical and #iochemical engineering since
large scale cultivation o, microorganisms and cells- their
do+nstream $rocessing are #ased on them. It c,0es t, us as a
great blessing...
Biotechnology utilies the techni3ue called genetic engineering or
recom#inant GA technology +here a microorganism is isolated its
genetic material is cut- mani$ulated- sealed- again inserted in an
organism and allo+ed to gro+ in a suita#le environment under
controlled conditions to get the desired $roduct. It loo*s easy #ut is a
very tedious jo# and it ta*es years ,or a research to achieve its goal.
i*e every other thing- bi,techn,l,g- t,, has s,0e
har0/ul i0pacts"
7. enetic engineering is a very vital $art o, #iotechnology and
the cost o, trans,erring genes ,rom one s$ecies to another is
very e$ensive- +hich re3uires a huge amount o, ca$ital
investment.
$he c,st ,/ pr,ducing geneticall-> 0,di2ed plants and
ani0als are s7-> r,c7eting and the duration o, return are also
not $redicta#le.
5. enetic engineering crosses #oundaries o, re$roduction #y
crossing genes o, s$ecies that are com$letely unrelated hence
giving rise to haardous results as +ell as also increasing the ris*
o, harming multi$le s$ecies.
L. =hen genetic material ,rom certain viruses is used in the
$roduction o, transgenic cro$s- there are chances that these virus
genes +ill com#ine +ith cro$ genes to $roduce more destructive
viruses. The consum$tion o, such cro$s is haardous to human
health and can cause several li,e9 threatening ailments. It can also
result in cancer- o,ten malignant as +ell.
H. Biotechnology also $oses a num#er o, environmental threats.
enetically modi4es cro$s o,ten in,ect monarch #utteries and
other insect s$ecies.

 The a$$lications o, #iotechnology are so #road- and the advantages so

com$elling- that virtually every industry is using this technology.
evelo$ments are under+ay in areas as diverse as $harmaceuticals-
diagnostics- tetiles- a3uaculture- ,orestry- chemicals-
household
$roducts- environmental cleanu$- ,ood $rocessing and ,orensics to name
a ,e+. Biotechnology is ena#ling these industries to ma*e ne+ or #etter
$roducts- o,ten +ith greater s$eed- e>ciency and Nei#ility.
Bi,techn,l,g- 0ust c,ntinue t, be care/ull- regulated s, that
the 0a3i0u0 bene2ts are recei1ed 5ith the least ris7. 

Bi#liogra$hy
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http?@@555.gene5atch.,rg@sub>:;: http?
@@en.5i7ipedia.,rg@hu0ulin http?
@@555.bi,techarticles.c,0@'thers>Article@Hu0an>

Insulin>and>#ec,0binant>!NA>$echn,l,g->D=.ht0l https?
@@isaaa.,rg@res,urces@publicati,ns@p,c7et7@<@de/ault.

asp http?
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https?@@en.5i7ipedia.,rg@5i7i@GeneEtherap- https?
@@en.5i7ipedia.,rg@5i7i@Aden,sineEdea0inaseEde2cie

nc- http?
@@555.diabetes.c,.u7@insulin@ani0al>insulin.ht0l
Bi,l,g- te3tb,,7 )N.C.".#.$* Class 8;th

Contents

• Introduction
• (istory
• Biotechnology in Agriculture
• enetically Modi4ed Cro$s
• @GA Inter,erence '@GAi)
 • Bt toin
• Bt cotton
• Biotechnology in Medicine
• enetically engineered
insulin '(umulin)
• ene thera$y
• Conclusion
• Bi#liogra$hy