UNIVERSIDAD DISTRITAL FRANCISCO JOS DE CALDAS FACULTAD DE CIENCIAS DE LA EDUCACIN LIC.

EN BIOLOGA GENTICA CELULAR Y MOLECULAR Cristhel Moreno Gutirrez COD: 20072140068

DISEO DE PRIMERS
SIMPLE: Planta: Solanum lycopersicum

Solanum lycopersicum beta-galactosidase (TBG2), mRNA NCBI Reference Sequence: NM_001247369.1 FASTAGraphics
nuccore 1 genbank

Go to: LOCUS DEFINITION ACCESSION VERSION KEYWORDS SOURCE

NM_001247369 3001 bp mRNA linear PLN 23-JAN-2012 Solanum lycopersicum beta-galactosidase (TBG2), mRNA. NM_001247369 NM_001247369.1 GI:350537548 . Solanum lycopersicum (Lycopersicon esculentum)

Solanum lycopersicum Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliophyta; eudicotyledons; core eudicotyledons; asterids; lamiids; Solanales; Solanaceae; Solanoideae; Solaneae; Solanum; Lycopersicon. REFERENCE 1 (bases 1 to 3001) AUTHORS Smith,D.L. and Gross,K.C. TITLE A family of at least seven beta-galactosidase genes is expressed during tomato fruit development JOURNAL Plant Physiol. 123 (3), 1173-1183 (2000) PUBMED 10889266 COMMENT PROVISIONAL REFSEQ: This record has not yet been subject to final NCBI review. The reference sequence was derived from AF154420.1. FEATURES Location/Qualifiers source 1..3001 /organism="Solanum lycopersicum" /mol_type="mRNA" /db_xref="taxon:4081" /chromosome="9" /map="IL 9-3" gene 1..3001 /gene="TBG2" /note="beta-galactosidase" /db_xref="GeneID:543608" misc_feature 14..16 /gene="TBG2" /note="upstream in-frame stop codon" CDS 23..2701 /gene="TBG2" /EC_number="3.2.1.23" /function="beta-galactosidase activity as measured by release of galactose from synthetic substrates such as p-nitrophenyl-beta-D-galactopyranoside" /note="targeted to the plasma membrane" /codon_start=1 /product="beta-galactosidase precursor" /protein_id="NP_001234298.1" /db_xref="GI:350537549" /db_xref="GeneID:543608" /translation="MVEAMSRRKTLNFPLILTVLTIHFVIVAGEYFKPFNVTYDNRAL IIGGKRRMLISAGIHYPRATPEMWPTLIARSKEGGADVIETYTFWNGHEPTRGQYNFE GRYDIVKFAKLVGSHGLFLFIRIGPYACAEWNFGGFPIWLRDIPGIEFRTDNAPFKEE MERYVKKIVDLMISESLFSWQGGPIILLQIENEYGNVESSFGPKGKLYMKWAAEMAVG LGAGVPWVMCRQTDAPEYIIDTCNAYYCDGFTPNSEKKPKIWTENWNGWFADWGERLP YRPSEDIAFAIARFFQRGGSLQNYYMYFGGTNFGRTAGGPTQITSYDYDAPLDEYGLL RQPKWGHLKDLHAAIKLCEPALVAADSPQYIKLGPKQEAHVYRGTSNNIGQYMSLNEG ICAAFIANIDEHESATVKFYGQEFTLPPWSVVFCQIAEIQLSTQLRWGHKLQSKQWAQ ILFQLGIILCFYKLSLKASSESFSQSWMTLKEPLGVWGDKNFTSKGILEHLNVTKDQS DYLWYLTRIYISDDDISFWEENDVSPTIDIDSMRDFVRIFVNGQLAGSVKGKWIKVVQ PVKLVQGYNDILLLSETVGLQNYGAFLEKDGAGFKGQIKLTGCKSGDINLTTSLWTYQ VGLRGEFLEVYDVNSTESAGWTEFPTGTTPSVFSWYKTKFDAPGGTDPVALDFSSMGK GQAWVNGHHVGRYWTLVAPNNGCGRTCDYRGAYHSDKCRTNCGEITQAWYHIPRSWLK TLNNVLVIFEETDKTPFDISISTRSTETICAQVSEKHYPPLHKWSHSEFDRKLSLMDK TPEMHLQCDEGHTISSIEFASYGSPNGSCQKFSQGKCHAANSLSVVSQACIGRTSCSI GISNGVFGDPCRHVVKSLAVQAKCSPPPDLSTSASS" sig_peptide 23..109 /gene="TBG2"

ORGANISM

ORIGIN 1 61 121 181 241 301 361 421 481 541 601 661 721 781 841 901 961 1021 1081 1141 1201 1261 1321 1381 1441 1501 1561 1621 1681 1741 1801 1861 1921 1981 2041 2101 2161 2221 2281 2341 2401 2461 2521 2581 2641 2701 2761 2821 2881 2941 3001 // atccattttc tccgttaata gccgttcaat tatctccgcc taggagcaaa gccaaccagg agtcggatct gaacttcggg taatgcacca atctgagtcg atatggaaat tgaaatggct tccagaatac cgagaagaaa aagacttcca tgggggcagc tggtggccca actacgtcaa accagctctt acatgtctat atgcgcagca tcaagagttc ttcaacacag tcagttggga ttcacaatct ttctaaagga tctgaccagg tccaacaatt tgcaggtagt atacaacgac ggagaaggat tatcaatctc agtatatgat tccgtcagtc tgctcttgat aagatattgg tgcttaccac tatacctaga taaaactccg atcggaaaag gtctctgatg ttctattgaa aaaatgccat ttgcagcatt tttggctgtt aggagactct tcccctgccc aaaacgtaca ataatgttgc aaactcctgt t atctgagaca ctaacggtgt gtcacctacg ggaattcact gaaggtggtg ggacagtaca catggactgt ggattcccca ttcaaggagg ctcttttcgt gttgaaagct gttggtcttg atcatagata ccgaaaattt tatagacctt ttacagaact actcaaatca cctaaatggg gttgctgctg cgtggaacat tttattgcaa actttacctc ctaaggtggg ataattcttt tggatgacat atactggagc atatatattt gatattgata gtgaaaggca atactgctat ggggcaggtt acaacatctt gtcaatagta ttttcgtggt tttagtagca actttggttg tctgataaat tcatggctaa tttgatattt cactatccac gataaaacac tttgcaagct gctgcaaatt ggcatttcca caagcaaaat ggtaacacgt ccgagccctc tggacgattt ttatgcatat ctgtcaaaga aaatggtaga taactatcca ataaccgagc accctcgcgc cagatgtcat attttgaagg tcctctttat tatggcttcg agatggagcg ggcaaggtgg cattcggtcc gtgctggtgt cttgtaatgc ggactgagaa ccgaggatat attatatgta ctagctatga gccatttgaa attcacctca ccaacaacat atattgatga catggtcagt ggcacaaact gtttctacaa tgaaggagcc atctgaatgt ctgatgatga gcatgcgtga aatggatcaa tatctgagac ttaaaggtca tatggaccta ctgaaagtgc acaagacaaa tgggaaaagg caccaaataa gtaggacaaa agacattaaa ccatttctac ctctacataa cagaaatgca atggaagtcc ccttgtctgt atggtgtatt gctcaccacc taacctttta tgctacattt tacttgtaaa gagctgcaaa attttaacaa agctatgagc ctttgtgatc tctcatcatc cactcctgag cgagacttat aagatatgat tcgaataggt tgatatacct ctatgttaaa tcctatcatt caaggggaag tccatgggtc atactattgt ttggaatgga tgcatttgca ttttggtggg ttatgatgct ggatctgcat gtatattaaa tggccaatat acatgaatca ggtattctgc tcaatcaaaa gttatcacta acttggtgtg gacaaaagac catctcattt ttttgttcgc ggtggttcaa ggtgggattg gataaagctt ccaggtgggg aggatggact gtttgatgcc tcaggcatgg tggatgtgga ctgtggagag taatgtacta gcgttctact gtggtctcat cttgcagtgt gaatggcagc tgtatctcag tggagatcca accagacctc gaacgaaacg ctcagatcgc tatttggtta ttatatgaca caccatttat agaagaaaaa gtcgccggcg ggcggtaaac atgtggccca acattttgga attgtcaagt ccttatgcct ggaatagaat aagatagttg ttgctgcaga ttatatatga atgtgcaggc gatgggttca tggtttgcag attgctcgtt acaaattttg ccactggatg gctgctataa ctgggaccaa atgtccttaa gcaacagtga cagattgcag cagtgggctc aaagcaagct tggggtgaca cagtctgatt tgggaggaaa atttttgtta cctgttaagc cagaattatg acaggatgca cttagaggcg gagtttccca ccaggcggga gttaatggcc agaacttgtg attactcagg gttatctttg gaaaccattt tcggagtttg gacgaaggac tgtcaaaagt gcttgtatag tgtcgacacg agcacttcag atcccttaaa atcgttacaa ctgtatataa aagtaacaaa taaaagttag cactgaattt agtatttcaa gccgtatgct cattgatagc atggtcatga tcgcaaagct gtgcagaatg ttcgaacaga atcttatgat ttgaaaatga aatgggctgc aaactgatgc cgccgaattc attggggtga tctttcaacg gccggactgc aatatggact agctttgtga aacaggaggc atgaaggcat aattttacgg aaatacagct agattctgtt cggaaagttt agaatttcac acctgtggta atgatgttag atgggcagct tggttcaggg gtgccttctt aaagcgggga aattcctgga ctggtacaac cagatccagt accatgtagg attatcgtgg cctggtacca aagaaacaga gtgctcaagt acagaaagtt atacaatctc tctcacaagg gaagaactag ttgtgaagag cttcctcgtg gtccactcgt tcaggcggag aatgaaagga tgaaaataga ttaacatgat

PRIMER REALIZADO EN PRIMER 3:

Primer3 Output

No mispriming library specified Using 1-based sequence positions OLIGO start len tm LEFT PRIMER 356 20 56.95 RIGHT PRIMER 508 20 57.23 SEQUENCE SIZE: 3001 INCLUDED REGION SIZE: 3001

gc% 50.00 50.00

any 4.00 3.00

3' seq 2.00 AAGCTAGTCGGATCTCATGG 3.00 CTCCATCTCCTCCTTGAATG

PRODUCT SIZE: 153, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 3.00 1 ATCCATTTTCATCTGAGACAAAATGGTAGAAGCTATGAGCAGAAGAAAAACACTGAATTT 61 TCCGTTAATACTAACGGTGTTAACTATCCACTTTGTGATCGTCGCCGGCGAGTATTTCAA 121 GCCGTTCAATGTCACCTACGATAACCGAGCTCTCATCATCGGCGGTAAACGCCGTATGCT 181 TATCTCCGCCGGAATTCACTACCCTCGCGCCACTCCTGAGATGTGGCCCACATTGATAGC 241 TAGGAGCAAAGAAGGTGGTGCAGATGTCATCGAGACTTATACATTTTGGAATGGTCATGA

301 GCCAACCAGGGGACAGTACAATTTTGAAGGAAGATATGATATTGTCAAGTTCGCAAAGCT >>>>> 361 AGTCGGATCTCATGGACTGTTCCTCTTTATTCGAATAGGTCCTTATGCCTGTGCAGAATG >>>>>>>>>>>>>>> 421 GAACTTCGGGGGATTCCCCATATGGCTTCGTGATATACCTGGAATAGAATTTCGAACAGA 481 TAATGCACCATTCAAGGAGGAGATGGAGCGCTATGTTAAAAAGATAGTTGATCTTATGAT <<<<<<<<<<<<<<<<<<<< 541 ATCTGAGTCGCTCTTTTCGTGGCAAGGTGGTCCTATCATTTTGCTGCAGATTGAAAATGA 601 ATATGGAAATGTTGAAAGCTCATTCGGTCCCAAGGGGAAGTTATATATGAAATGGGCTGC 661 TGAAATGGCTGTTGGTCTTGGTGCTGGTGTTCCATGGGTCATGTGCAGGCAAACTGATGC 721 TCCAGAATACATCATAGATACTTGTAATGCATACTATTGTGATGGGTTCACGCCGAATTC 781 CGAGAAGAAACCGAAAATTTGGACTGAGAATTGGAATGGATGGTTTGCAGATTGGGGTGA 841 AAGACTTCCATATAGACCTTCCGAGGATATTGCATTTGCAATTGCTCGTTTCTTTCAACG 901 TGGGGGCAGCTTACAGAACTATTATATGTATTTTGGTGGGACAAATTTTGGCCGGACTGC 961 TGGTGGCCCAACTCAAATCACTAGCTATGATTATGATGCTCCACTGGATGAATATGGACT 1021 ACTACGTCAACCTAAATGGGGCCATTTGAAGGATCTGCATGCTGCTATAAAGCTTTGTGA 1081 ACCAGCTCTTGTTGCTGCTGATTCACCTCAGTATATTAAACTGGGACCAAAACAGGAGGC 1141 ACATGTCTATCGTGGAACATCCAACAACATTGGCCAATATATGTCCTTAAATGAAGGCAT 1201 ATGCGCAGCATTTATTGCAAATATTGATGAACATGAATCAGCAACAGTGAAATTTTACGG 1261 TCAAGAGTTCACTTTACCTCCATGGTCAGTGGTATTCTGCCAGATTGCAGAAATACAGCT 1321 TTCAACACAGCTAAGGTGGGGGCACAAACTTCAATCAAAACAGTGGGCTCAGATTCTGTT 1381 TCAGTTGGGAATAATTCTTTGTTTCTACAAGTTATCACTAAAAGCAAGCTCGGAAAGTTT 1441 TTCACAATCTTGGATGACATTGAAGGAGCCACTTGGTGTGTGGGGTGACAAGAATTTCAC 1501 TTCTAAAGGAATACTGGAGCATCTGAATGTGACAAAAGACCAGTCTGATTACCTGTGGTA

Las bases nitrogenadas que estn en rojo son las complementarias al primers LEFT y RIGHT. Y la zona verde corresponde al amplicon.

1561 TCTGACCAGGATATATATTTCTGATGATGACATCTCATTTTGGGAGGAAAATGATGTTAG 1621 TCCAACAATTGATATTGATAGCATGCGTGATTTTGTTCGCATTTTTGTTAATGGGCAGCT 1681 TGCAGGTAGTGTGAAAGGCAAATGGATCAAGGTGGTTCAACCTGTTAAGCTGGTTCAGGG 1741 ATACAACGACATACTGCTATTATCTGAGACGGTGGGATTGCAGAATTATGGTGCCTTCTT 1801 GGAGAAGGATGGGGCAGGTTTTAAAGGTCAGATAAAGCTTACAGGATGCAAAAGCGGGGA 1861 TATCAATCTCACAACATCTTTATGGACCTACCAGGTGGGGCTTAGAGGCGAATTCCTGGA 1921 AGTATATGATGTCAATAGTACTGAAAGTGCAGGATGGACTGAGTTTCCCACTGGTACAAC 1981 TCCGTCAGTCTTTTCGTGGTACAAGACAAAGTTTGATGCCCCAGGCGGGACAGATCCAGT 2041 TGCTCTTGATTTTAGTAGCATGGGAAAAGGTCAGGCATGGGTTAATGGCCACCATGTAGG 2101 AAGATATTGGACTTTGGTTGCACCAAATAATGGATGTGGAAGAACTTGTGATTATCGTGG 2161 TGCTTACCACTCTGATAAATGTAGGACAAACTGTGGAGAGATTACTCAGGCCTGGTACCA 2221 TATACCTAGATCATGGCTAAAGACATTAAATAATGTACTAGTTATCTTTGAAGAAACAGA 2281 TAAAACTCCGTTTGATATTTCCATTTCTACGCGTTCTACTGAAACCATTTGTGCTCAAGT 2341 ATCGGAAAAGCACTATCCACCTCTACATAAGTGGTCTCATTCGGAGTTTGACAGAAAGTT 2401 GTCTCTGATGGATAAAACACCAGAAATGCACTTGCAGTGTGACGAAGGACATACAATCTC 2461 TTCTATTGAATTTGCAAGCTATGGAAGTCCGAATGGCAGCTGTCAAAAGTTCTCACAAGG 2521 AAAATGCCATGCTGCAAATTCCTTGTCTGTTGTATCTCAGGCTTGTATAGGAAGAACTAG 2581 TTGCAGCATTGGCATTTCCAATGGTGTATTTGGAGATCCATGTCGACACGTTGTGAAGAG 2641 TTTGGCTGTTCAAGCAAAATGCTCACCACCACCAGACCTCAGCACTTCAGCTTCCTCGTG 2701 AGGAGACTCTGGTAACACGTTAACCTTTTAGAACGAAACGATCCCTTAAAGTCCACTCGT 2761 TCCCCTGCCCCCGAGCCCTCTGCTACATTTCTCAGATCGCATCGTTACAATCAGGCGGAG

2821 AAAACGTACATGGACGATTTTACTTGTAAATATTTGGTTACTGTATATAAAATGAAAGGA 2881 ATAATGTTGCTTATGCATATGAGCTGCAAATTATATGACAAAGTAACAAATGAAAATAGA 2941 AAACTCCTGTCTGTCAAAGAATTTTAACAACACCATTTATTAAAAGTTAGTTAACATGAT 3001 T KEYS (in order of precedence): >>>>>> left primer <<<<<< right primer ADDITIONAL OLIGOS start len tm gc% any 3' seq 1 LEFT PRIMER 1720 20 56.93 50.00 6.00 2.00 ACCTGTTAAGCTGGTTCAGG RIGHT PRIMER 1964 20 57.28 50.00 5.00 3.00 ACTCAGTCCATCCTGCACTT PRODUCT SIZE: 245, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 3.00 2 LEFT PRIMER 138 20 56.95 50.00 6.00 2.00 ACGATAACCGAGCTCTCATC RIGHT PRIMER 322 20 57.36 50.00 4.00 0.00 ATTGTACTGTCCCCTGGTTG PRODUCT SIZE: 185, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00 3 LEFT PRIMER 1720 20 56.93 50.00 6.00 2.00 ACCTGTTAAGCTGGTTCAGG RIGHT PRIMER 1898 20 57.40 50.00 5.00 0.00 CCACCTGGTAGGTCCATAAA PRODUCT SIZE: 179, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00 4 LEFT PRIMER 358 20 57.25 55.00 4.00 2.00 GCTAGTCGGATCTCATGGAC RIGHT PRIMER 508 20 57.23 50.00 3.00 3.00 CTCCATCTCCTCCTTGAATG PRODUCT SIZE: 151, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 3.00 Statistics con too in in no tm tm high high sid many tar excl bad GC too too any 3' poly ered Ns get reg GC% clamp low high compl compl X Left 14002 0 0 0 10918 0 362 1800 7 18 0 Right 13977 0 0 0 10906 0 361 1807 2 6 0 Pair Stats: considered 1928, unacceptable product size 1901, high end compl 7, ok 20 primer3 release 1.1.4 (primer3_results.cgi release 0.4.0) high end stab 143 154

ok 754 741

PRIMER REALIZADO BIBISERV:

DEGENERADO:

Dentro de estas opciones no se encuentra una especie que pertenezca a la misma familia (SOLANACEAE), para hacer el primer GRUPAL, pero en este caso el primer DEGENERADO lo podemos realizar con cualquiera de estas opciones (menos la primera), puesto que no es de la misma especie.