请问知道基因组DNA序列如何设计引物
丁香园论坛
7007
这是在NCBI中查到的大豆贮藏蛋白基因Gy1的DNA序列，如果我想扩这个基因做表达用。如何扩，里面含有内含子，是不是把CDS连起来进行设计引物，后进行RT－PCR，还是直接扩增基因组DNA好。我是新手，请高手指教。
LOCUS GMGY1 3527 bp DNA linear PLN 10-FEB-1999
DEFINITION Soybean Gy1 gene for glycinin subunit G1.
ACCESSION X15121
VERSION X15121.1 GI:18634
KEYWORDS glycinin; Gy1 gene.
SOURCE Glycine max (soybean)
ORGANISM Glycine max
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; eudicotyledons; core eudicots;
rosids; eurosids I; Fabales; Fabaceae; Papilionoideae; Phaseoleae;
Glycine.
REFERENCE 1 (bases 1 to 3527)
AUTHORS Sims,T.L. and Goldberg,R.B.
TITLE The glycinin Gy1 gene from soybean
JOURNAL Nucleic Acids Res. 17 (11), 4386 (1989)
MEDLINE 89296498
PUBMED 2740229
REFERENCE 2 (bases 1 to 3527)
AUTHORS Nielsen,N.C.
TITLE Direct Submission
JOURNAL Submitted (26-APR-1989) Nielsen N.C., USDA-ARS Purdue University,
Agronomy Dept, West Lafayette IN 47907, U S A
FEATURES Location/Qualifiers
source 1..3527
/organism="Glycine max"
/mol_type="genomic DNA"
/strain="var. Dare"
/db_xref="taxon:3847"
/clone="lambda A28-30"
/tissue_type="leaf."
/clone_lib="Alu1/Hae3 linker"
misc_feature 211..219
/note="caca box"
misc_feature 386..393
/note="catgcatg element"
misc_feature 522..549
/note="legumin box"
misc_feature 530..537
/note="catgcatg element"
TATA_signal 607..612
precursor_RNA 639..3282
/note="Gy1 transcript"
exon 639..976
/number=1
CDS join(691..976,1205..1458,1750..2307,2689..3078)
/codon_start=1
/product="glycinin subunit G1"
/protein_id="CAA33215.1"
/db_xref="GI:18635"
/db_xref="GOA:P04776"
/db_xref="Swiss-Prot:P04776"
/translation="MAKLVFSLCFLLFSGCCFAFSSREQPQQNECQIQKLNALKPDNR
IESEGGLIETWNPNNKPFQCAGVALSRCTLNRNALRRPSYTNGPQEIYIQQGKGIFGM
IYPGCPSTFEEPQQPQQRGQSSRPQDRHQKIYNFREGDLIAVPTGVAWWMYNNEDTPV
VAVSIIDTNSLENQLDQMPRRFYLAGNQEQEFLKYQQEQGGHQSQKGKHQQEEENEGG
SILSGFTLEFLEHAFSVDKQIAKNLQGENEGEDKGAIVTVKGGLSVIKPPTDEQQQRP
QEEEEEEEDEKPQCKGKDKHCQRPRGSQSKSRRNGIDETICTMRLRHNIGQTSSPDIY
NPQAGSVTTATSLDFPALSWLRLSAEFGSLRKNAMFVPHYNLNANSIIYALNGRALIQ
VVNCNGERVFDGELQEGRVLIVPQNFVVAARSQSDNFEYVSFKTNDTPMIGTLAGANS
LLNALPEEVIQHTFNLKSQQARQIKNNNPFKFLVPPQESQKRAVA"
intron 977..1204
/number=1
exon 1205..1458
/number=2
intron 1459..1749
/number=2
exon 1750..2307
/number=3
intron 2308..2688
/number=3
exon 2689..3282
/number=4
polyA_signal 3136..3141
polyA_signal 3250..3255
ORIGIN
1 tagcctaagt acgtactcaa aatgccaaca aataaaaaaa aagttgcttt aataatgcca
61 aaacaaatta ataaaacact tacaacaccg gatttttttt aattaaaatg tgccatttag
121 gataaatagt taatattttt aataattatt taaaaagccg tatctactaa aatgattttt
181 atttggttga aaatattaat atgtttaaat caacacaatc tatcaaaatt aaactaaaaa
241 aaaaataagt gtacgtggtt aacattagta cagtaatata agaggaaaat gagaaattaa
301 gaaattgaaa gcgagtctaa tttttaaatt atgaacctgc atatataaaa ggaaagaaag
361 aatccaggaa gaaaagaaat gaaaccatgc atggtcccct cgtcatcacg agtttctgcc
421 atttgcaata gaaacactga aacacctttc tctttgtcac ttaattgaga tgccgaagcc
481 acctcacacc atgaacttca tgaggtgtag cacccaaggc ttccatagcc atgcatactg
541 aagaatgtct caagctcagc accctacttc tgtgacgttg tccctcattc accttcctct
601 cttccctata aataaccacg cctcaggttc tccgcttcac aactcaaaca ttctcctcca
661 ttggtcctta aacactcatc agtcatcacc atggccaagc tagttttttc cctttgtttt
721 ctgcttttca gtggctgctg cttcgctttc agttccagag agcagcctca gcaaaacgag
781 tgccagatcc aaaaactcaa tgccctcaaa ccggataacc gtatagagtc agaaggaggg
841 ctcattgaga catggaaccc taacaacaag ccattccagt gtgccggtgt tgccctctct
901 cgctgcaccc tcaaccgcaa cgcccttcgt agaccttcct acaccaacgg tccccaggaa
961 atctacatcc aacaaggtcc atcttgtcca aacttcacat ataaatatat aatagactta
1021 aatatgttta agggtttgat aaatgaggga attttatttt agatttttaa taattttttt
1081 tgttttgagt ttttatatat taaaattttt gttttgattt cttccatatg acgtaacata
1141 atcatatcat tgataatgtt gggttcctaa tttttgtttg tttgttgttt tgtaatatga
1201 ataggtaagg gtatttttgg catgatatac ccgggttgtc ctagcacatt tgaagagcct
1261 caacaacctc aacaaagagg acaaagcagc agaccacaag accgtcacca gaagatctat
1321 aacttcagag agggtgattt gatcgcagtg cctactggtg ttgcatggtg gatgtacaac
1381 aatgaagaca ctcctgttgt tgccgtttct attattgaca ccaacagctt ggagaaccag
1441 ctcgaccaga tgcctagggt gagccacata gcaatattag atattataat tctttaaagg
1501 tttaaatatc attttagttc gtggagttgc actttctaat ttagtaccta tagattaaaa
1561 tatgccaatt gaatccttat agttgtgttt ttttatccaa tttggttctt gtcttgaaat
1621 aaatggacaa tattgtagct gataaaaaaa ggaaactgga ctacattgta acgttaagat
1681 tagaattctt aagttctaat actagctggt tacagattga caactatttg ttttgacaat
1741 tcttggcaga gattctatct tgctgggaac caagagcaag agtttctaaa atatcagcaa
1801 gagcaaggag gtcatcaaag ccagaaagga aagcatcagc aagaagaaga aaacgaagga
1861 ggcagcatat tgagtggctt caccctggaa ttcttggaac atgcattcag cgtggacaag
1921 cagatagcga aaaacctaca aggagagaac gaaggggaag acaagggagc cattgtgaca
1981 gtgaaaggag gtctgagcgt gataaaacca cccacggacg agcagcaaca aagaccccag
2041 gaagaggaag aagaagaaga ggatgagaag ccacagtgca agggtaaaga caaacactgc
2101 caacgccccc gaggaagcca aagcaaaagc agaagaaatg gcattgacga gaccatatgc
2161 accatgagac ttcgccacaa cattggccag acttcatcac ctgacatcta caaccctcaa
2221 gccggtagcg tcacaaccgc caccagcctt gacttcccag ccctctcgtg gctcagactc
2281 agtgctgagt ttggatctct ccgcaaggta cgtacatcat tcatcaaaga tcaacataca
2341 tttatacatt aaactaatat ttgttgccaa atatttatta attttattga taattaattt
2401 ttttagaaaa tttgtttgat cacttttaat ggagtctttc atcttaatta cattatttat
2461 acttagacta atgatttatt gattaataat aatcttagat acactataaa atgtgtgacg
2521 gagttatctt aacacttgca tggattctat cttttctgtc tttatatata gaaatagaga
2581 gaaaaaaaaa gaaaagattg atgaaaaaag caaaacaaaa aatagtatta ttataaaaat
2641 attggatgaa tttgttgtga ctcttgcatg cattgatgta cgatgcagaa tgcaatgttc
2701 gtgccacact acaacctgaa cgcgaacagc ataatatacg cattgaatgg acgggcattg
2761 atacaagtgg tgaattgcaa cggtgagaga gtgtttgatg gagagctgca agagggacgg
2821 gtgctgatcg tgccacaaaa ctttgtggtg gctgcaagat cacagagtga caacttcgag
2881 tatgtgtcat tcaagaccaa tgatacaccc atgatcggca ctcttgcagg ggcaaactca
2941 ttgttgaacg cattaccaga ggaagtgatt cagcacactt tcaacctaaa aagccagcag
3001 gccaggcaga taaagaacaa caaccctttc aagttcctgg ttccacctca ggagtctcag
3061 aagagagctg tggcttagag ccctttttgt atgtgctacc ccacttttgt ctttttggca
3121 atagtgctag caaccaataa ataataataa taataatgaa taagaaaaca aaggctttag
3181 cttgcctttt gttcactgta aaataataat gtaagtactc tctataatga gtcacgaaac
3241 ttttgcggga ataaaaggag aaattccaat gagttttctg tcaaatcttc ttttgtctct
3301 ctctctctct cttttttttt tctttcttct gagcttcttg caaaacaaaa ggcaaacaat
3361 aacgattggt ccaatgatag ttagcttgat cgatgatatc tttaggaagt gttggcagga
3421 caggacatga tgtagaagac taaaattgaa agtattgcag acccaatagt tgaagattaa
3481 ctttaagaat gaagacgtct tatcaggttc ttcatgactt ggagctc
LOCUS GMGY1 3527 bp DNA linear PLN 10-FEB-1999
DEFINITION Soybean Gy1 gene for glycinin subunit G1.
ACCESSION X15121
VERSION X15121.1 GI:18634
KEYWORDS glycinin; Gy1 gene.
SOURCE Glycine max (soybean)
ORGANISM Glycine max
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; eudicotyledons; core eudicots;
rosids; eurosids I; Fabales; Fabaceae; Papilionoideae; Phaseoleae;
Glycine.
REFERENCE 1 (bases 1 to 3527)
AUTHORS Sims,T.L. and Goldberg,R.B.
TITLE The glycinin Gy1 gene from soybean
JOURNAL Nucleic Acids Res. 17 (11), 4386 (1989)
MEDLINE 89296498
PUBMED 2740229
REFERENCE 2 (bases 1 to 3527)
AUTHORS Nielsen,N.C.
TITLE Direct Submission
JOURNAL Submitted (26-APR-1989) Nielsen N.C., USDA-ARS Purdue University,
Agronomy Dept, West Lafayette IN 47907, U S A
FEATURES Location/Qualifiers
source 1..3527
/organism="Glycine max"
/mol_type="genomic DNA"
/strain="var. Dare"
/db_xref="taxon:3847"
/clone="lambda A28-30"
/tissue_type="leaf."
/clone_lib="Alu1/Hae3 linker"
misc_feature 211..219
/note="caca box"
misc_feature 386..393
/note="catgcatg element"
misc_feature 522..549
/note="legumin box"
misc_feature 530..537
/note="catgcatg element"
TATA_signal 607..612
precursor_RNA 639..3282
/note="Gy1 transcript"
exon 639..976
/number=1
CDS join(691..976,1205..1458,1750..2307,2689..3078)
/codon_start=1
/product="glycinin subunit G1"
/protein_id="CAA33215.1"
/db_xref="GI:18635"
/db_xref="GOA:P04776"
/db_xref="Swiss-Prot:P04776"
/translation="MAKLVFSLCFLLFSGCCFAFSSREQPQQNECQIQKLNALKPDNR
IESEGGLIETWNPNNKPFQCAGVALSRCTLNRNALRRPSYTNGPQEIYIQQGKGIFGM
IYPGCPSTFEEPQQPQQRGQSSRPQDRHQKIYNFREGDLIAVPTGVAWWMYNNEDTPV
VAVSIIDTNSLENQLDQMPRRFYLAGNQEQEFLKYQQEQGGHQSQKGKHQQEEENEGG
SILSGFTLEFLEHAFSVDKQIAKNLQGENEGEDKGAIVTVKGGLSVIKPPTDEQQQRP
QEEEEEEEDEKPQCKGKDKHCQRPRGSQSKSRRNGIDETICTMRLRHNIGQTSSPDIY
NPQAGSVTTATSLDFPALSWLRLSAEFGSLRKNAMFVPHYNLNANSIIYALNGRALIQ
VVNCNGERVFDGELQEGRVLIVPQNFVVAARSQSDNFEYVSFKTNDTPMIGTLAGANS
LLNALPEEVIQHTFNLKSQQARQIKNNNPFKFLVPPQESQKRAVA"
intron 977..1204
/number=1
exon 1205..1458
/number=2
intron 1459..1749
/number=2
exon 1750..2307
/number=3
intron 2308..2688
/number=3
exon 2689..3282
/number=4
polyA_signal 3136..3141
polyA_signal 3250..3255
ORIGIN
1 tagcctaagt acgtactcaa aatgccaaca aataaaaaaa aagttgcttt aataatgcca
61 aaacaaatta ataaaacact tacaacaccg gatttttttt aattaaaatg tgccatttag
121 gataaatagt taatattttt aataattatt taaaaagccg tatctactaa aatgattttt
181 atttggttga aaatattaat atgtttaaat caacacaatc tatcaaaatt aaactaaaaa
241 aaaaataagt gtacgtggtt aacattagta cagtaatata agaggaaaat gagaaattaa
301 gaaattgaaa gcgagtctaa tttttaaatt atgaacctgc atatataaaa ggaaagaaag
361 aatccaggaa gaaaagaaat gaaaccatgc atggtcccct cgtcatcacg agtttctgcc
421 atttgcaata gaaacactga aacacctttc tctttgtcac ttaattgaga tgccgaagcc
481 acctcacacc atgaacttca tgaggtgtag cacccaaggc ttccatagcc atgcatactg
541 aagaatgtct caagctcagc accctacttc tgtgacgttg tccctcattc accttcctct
601 cttccctata aataaccacg cctcaggttc tccgcttcac aactcaaaca ttctcctcca
661 ttggtcctta aacactcatc agtcatcacc atggccaagc tagttttttc cctttgtttt
721 ctgcttttca gtggctgctg cttcgctttc agttccagag agcagcctca gcaaaacgag
781 tgccagatcc aaaaactcaa tgccctcaaa ccggataacc gtatagagtc agaaggaggg
841 ctcattgaga catggaaccc taacaacaag ccattccagt gtgccggtgt tgccctctct
901 cgctgcaccc tcaaccgcaa cgcccttcgt agaccttcct acaccaacgg tccccaggaa
961 atctacatcc aacaaggtcc atcttgtcca aacttcacat ataaatatat aatagactta
1021 aatatgttta agggtttgat aaatgaggga attttatttt agatttttaa taattttttt
1081 tgttttgagt ttttatatat taaaattttt gttttgattt cttccatatg acgtaacata
1141 atcatatcat tgataatgtt gggttcctaa tttttgtttg tttgttgttt tgtaatatga
1201 ataggtaagg gtatttttgg catgatatac ccgggttgtc ctagcacatt tgaagagcct
1261 caacaacctc aacaaagagg acaaagcagc agaccacaag accgtcacca gaagatctat
1321 aacttcagag agggtgattt gatcgcagtg cctactggtg ttgcatggtg gatgtacaac
1381 aatgaagaca ctcctgttgt tgccgtttct attattgaca ccaacagctt ggagaaccag
1441 ctcgaccaga tgcctagggt gagccacata gcaatattag atattataat tctttaaagg
1501 tttaaatatc attttagttc gtggagttgc actttctaat ttagtaccta tagattaaaa
1561 tatgccaatt gaatccttat agttgtgttt ttttatccaa tttggttctt gtcttgaaat
1621 aaatggacaa tattgtagct gataaaaaaa ggaaactgga ctacattgta acgttaagat
1681 tagaattctt aagttctaat actagctggt tacagattga caactatttg ttttgacaat
1741 tcttggcaga gattctatct tgctgggaac caagagcaag agtttctaaa atatcagcaa
1801 gagcaaggag gtcatcaaag ccagaaagga aagcatcagc aagaagaaga aaacgaagga
1861 ggcagcatat tgagtggctt caccctggaa ttcttggaac atgcattcag cgtggacaag
1921 cagatagcga aaaacctaca aggagagaac gaaggggaag acaagggagc cattgtgaca
1981 gtgaaaggag gtctgagcgt gataaaacca cccacggacg agcagcaaca aagaccccag
2041 gaagaggaag aagaagaaga ggatgagaag ccacagtgca agggtaaaga caaacactgc
2101 caacgccccc gaggaagcca aagcaaaagc agaagaaatg gcattgacga gaccatatgc
2161 accatgagac ttcgccacaa cattggccag acttcatcac ctgacatcta caaccctcaa
2221 gccggtagcg tcacaaccgc caccagcctt gacttcccag ccctctcgtg gctcagactc
2281 agtgctgagt ttggatctct ccgcaaggta cgtacatcat tcatcaaaga tcaacataca
2341 tttatacatt aaactaatat ttgttgccaa atatttatta attttattga taattaattt
2401 ttttagaaaa tttgtttgat cacttttaat ggagtctttc atcttaatta cattatttat
2461 acttagacta atgatttatt gattaataat aatcttagat acactataaa atgtgtgacg
2521 gagttatctt aacacttgca tggattctat cttttctgtc tttatatata gaaatagaga
2581 gaaaaaaaaa gaaaagattg atgaaaaaag caaaacaaaa aatagtatta ttataaaaat
2641 attggatgaa tttgttgtga ctcttgcatg cattgatgta cgatgcagaa tgcaatgttc
2701 gtgccacact acaacctgaa cgcgaacagc ataatatacg cattgaatgg acgggcattg
2761 atacaagtgg tgaattgcaa cggtgagaga gtgtttgatg gagagctgca agagggacgg
2821 gtgctgatcg tgccacaaaa ctttgtggtg gctgcaagat cacagagtga caacttcgag
2881 tatgtgtcat tcaagaccaa tgatacaccc atgatcggca ctcttgcagg ggcaaactca
2941 ttgttgaacg cattaccaga ggaagtgatt cagcacactt tcaacctaaa aagccagcag
3001 gccaggcaga taaagaacaa caaccctttc aagttcctgg ttccacctca ggagtctcag
3061 aagagagctg tggcttagag ccctttttgt atgtgctacc ccacttttgt ctttttggca
3121 atagtgctag caaccaataa ataataataa taataatgaa taagaaaaca aaggctttag
3181 cttgcctttt gttcactgta aaataataat gtaagtactc tctataatga gtcacgaaac
3241 ttttgcggga ataaaaggag aaattccaat gagttttctg tcaaatcttc ttttgtctct
3301 ctctctctct cttttttttt tctttcttct gagcttcttg caaaacaaaa ggcaaacaat
3361 aacgattggt ccaatgatag ttagcttgat cgatgatatc tttaggaagt gttggcagga
3421 caggacatga tgtagaagac taaaattgaa agtattgcag acccaatagt tgaagattaa
3481 ctttaagaat gaagacgtct tatcaggttc ttcatgactt ggagctc
