| Summary: | 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.
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