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        请问知道基因组DNA序列如何设计引物

        丁香园论坛

        7480
        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
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