Analysis of human cytosolic NADP+ -dependent malic enzyme genomic structure.

• Main Authors: Fu-Hsin Chang, 張富信
• Other Authors: Wei-Yuan Chou
• Format: Others
• Language: zh-TW
• Published: 1996
• Online Access: http://ndltd.ncl.edu.tw/handle/38969128742953529510

碩士 === 國防醫學院 === 生物化學研究所 === 84 === A human genomic.lambda.bacteriophage library was screening for human malic enzyme gene [(S)-malate:NADP+ oxidoreductase; EC 1.1.1.40] were clones in the length of 15 kb for eath, B1, B46 and B93 were obtained using its cDNA as probes, the 5 kb genomic DNA fragment, BT487, was obtained by the polymerase chain reaction using human chromosome DNA as templates. These genomic DNA fragments consist of 9 exons and 10 introns. These genomic clones were analyzed by restriction mapping and Southern blot analysis. The B1 and B46 clones contain the 5' end of the malic enzyme cDNA and the B93 and BT487 clones contain the 3' end of the malic enzyme cDNA. The genomic fragments containing exon sequences and their adjacent regions were subcloned into the pBluescript vector and sequenced to locate the exon/intron boundaries. All these boundaries adhere to the GT/AG rule, they begin at the 5' end with the dinucleotide GT and terminate with the dinucleotide AG. The sizes of introns are determined by the polymerase chain reaction. Using the first exon and vector DNA sequence as primers, the 5'untranslated region in the length of 53 bps was obtained by the polymerase chain reaction.