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        请大家给我看看这对引物是不是有问题

        丁香园论坛

        1292
        我的课题涉及到CYP1A1的多态,从文献上(cancer res等级别的文章,而且重复的人挺多)查到它的一个MspI多态位点引物为5’-CAG TGA AGA GGT GTA GCC GCT-3'和5’-TAG GAG TCT TGT CTC ATG CCT-3’,但我有扩增出了其他引物的条件都没有能扩增出这对引物的目标带,做梯度从48到65度都没有带出来。下面是所查的该引物的Blast的资料和该基因这个位点的基因序列资料,请高手帮忙分析一下到底这对引物可不可以扩出来
        Blast资料:

        >gi|219566|dbj|D12525.1|HUMCYP1A12 Homo sapiens gene for cytochrome P450IA1, 3'flanking region
        Length = 650

        Score = 44.1 bits (22), Expect = 0.013
        Identities = 22/22 (100%)
        Strand = Plus / Minus


        Query: 21 ttaggagtcttgtctcatgcct 42
        ||||||||||||||||||||||
        Sbjct: 606 ttaggagtcttgtctcatgcct 585

        Score = 42.1 bits (21), Expect = 0.050
        Identities = 21/21 (100%)
        Strand = Plus / Plus


        Query: 1 cagtgaagaggtgtagccgct 21
        |||||||||||||||||||||
        Sbjct: 266 cagtgaagaggtgtagccgct 286

        >gi|13430063|gb|AF253322.1|AF253322 Homo sapiens cytochrome P450 (CYP1A1) and cytochrome P450 (CYP1A2)
        genes, complete cds
        Length = 43064

        Score = 44.1 bits (22), Expect = 0.013
        Identities = 22/22 (100%)
        Strand = Plus / Plus


        Query: 21 ttaggagtcttgtctcatgcct 42
        ||||||||||||||||||||||
        Sbjct: 4285 ttaggagtcttgtctcatgcct 4306

        Score = 42.1 bits (21), Expect = 0.050
        Identities = 21/21 (100%)
        Strand = Plus / Minus


        Query: 1 cagtgaagaggtgtagccgct 21
        |||||||||||||||||||||
        Sbjct: 4628 cagtgaagaggtgtagccgct 4608

        基因序列

        1: D12525. Homo sapiens gene...[gi:219566] Links

        LOCUS HUMCYP1A12 650 bp DNA linear PRI 13-FEB-2003
        DEFINITION Homo sapiens gene for cytochrome P450IA1, 3'flanking region.
        ACCESSION D12525 D01198
        VERSION D12525.1 GI:219566
        KEYWORDS Benzo(a)pyrene metabolism.
        SOURCE Homo sapiens (human)
        ORGANISM Homo sapiens
        Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;
        Mammalia; Eutheria; Primates; Catarrhini; Hominidae; Homo.
        REFERENCE 1
        AUTHORS Kawajiri,K., Watanabe,J., Gotoh,O., Tagashira,Y., Sogawa,K. and
        Fujii-Kuriyama,Y.
        TITLE Structure and drug inducibility of the human cytochrome P-450c gene
        JOURNAL Eur. J. Biochem. 159 (2), 219-225 (1986)
        MEDLINE 87004629
        PUBMED 3019683
        REFERENCE 2
        AUTHORS Kubota,M., Sogawa,K., Kaizu,Y., Sawaya,T., Watanabe,J.,
        Kawajiri,K., Gotoh,O. and Fujii-Kuriyama,Y.
        TITLE Xenobiotic responsive element in the 5'-upstream region of the
        human P-450c gene
        JOURNAL J. Biochem. 110 (2), 232-236 (1991)
        MEDLINE 92105053
        PUBMED 1761516
        REFERENCE 3 (bases 1 to 650)
        AUTHORS Hayashi,S., Watanabe,J., Nakachi,K. and Kawajiri,K.
        TITLE Genetic linkage of lung cancer-associated MspI polymorphisms with
        amino acid replacement in the heme binding region of the human
        cytochrome P450IA1 gene
        JOURNAL J. Biochem. 110 (3), 407-411 (1991)
        MEDLINE 92121159
        PUBMED 1722803
        REFERENCE 4 (bases 1 to 650)
        AUTHORS Hayashi,S.
        TITLE Direct Submission
        JOURNAL Submitted (04-OCT-1991) Shin-ichi Hayashi, Saitama Cancer Center
        Research Institute, Department of Biochemistry; 818 Komuro,
        Inamachi, Saitama 362, Japan (Tel:81-48-722-1111(ex.257),
        Fax:81-48-722-1739)
        FEATURES Location/Qualifiers
        source 1..650
        /organism="Homo sapiens"
        /mol_type="genomic DNA"
        /db_xref="taxon:9606"
        /chromosome="15"
        /map="15q22-qter"
        /clone="phP450mc-2"
        /cell_type="lymphocyte"
        /tissue_type="blood"
        /note="germline"
        polyA_signal 132..137
        /note="cytochrome P450IA1 (CYP1A1)"
        variation 400
        /note="Base substitution has occurred at this position in
        genotype C
        MspI polymorphism
        cancer susceptibility associated"
        /replace="c"
        ORIGIN
        1 acaatccttc tattctagcc tgcattgagc ttgcatgctt gcataagagc ttaagaaacc
        61 attgatttaa tgtaataggg aaaattctaa cccaggtatc caaaaatgtg taagaacaac
        121 tacctgagct aaataaagat attgttcaga aaatcctata ggtggagatt ttttgaatca
        181 taaatgattc atcactcgtc taaatactca ccctgaaccc cattctgtgt tgggttttac
        241 tgtagggagg aagaagagga ggtagcagtg aagaggtgta gccgctgcac ttaagcagtc
        301 tgtttgaggg acaagactct attttttgag acagggtccc caggtcatcc aggctggagt
        361 gcactggtac cattttgttt cactgtaacc tccacctcct gggctcacac gattctccca
        421 cctcagcctc tgagtagttg gggccgccag acgccaccac agcttttttt tttttttttt
        481 tttttttttg tagagatggg gtttcaccat gttgcccagg ctggtctcaa actcctgagc
        541 tcaagtgatc cacctgcctc agcctcccaa agtgctggga ttacaggcat gagacaagac
        601 tcctaatcac tgtgctgtct tagcgccctc tctaacttat cacaaattga
        //
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