聚合酶链反应(PCR)所得小麦高分子量谷蛋白亚基基因片段的克隆和顺序分析  

MOLECULAR CLONING AND SEQUENCING OF WHEAT HIGHMOLECULAR-WEIGHT GLUTENIN SUBUNIT GENE FRAGMENT USING POLYMERASE CHAIN REACTION

作　　者：李育庆 曹凯鸣[1] 忻骅[1] 顾其敏[1] 孙崇荣[1] 

机构地区：[1]复旦大学生物化学系

出　　处：《Acta Botanica Sinica》1992年第9期651-657,共7页Acta Botanica Sinica（植物学报：英文版）

基　　金：国家高技术研究发展计划(863计划)资助项目

摘　　要：应用聚合酶链反应(PCR)法,以两种人工合成的寡核苷酸片段为引物对,在Taq DNA聚合酶催化下,经35个反应循环后,从'扬麦4号'的核DNA中直接扩增出小麦(Triticumaestivum L.)高分子量(HMW)谷蛋白亚基的基因片段。体外扩增得到的片段克隆于pBSSK^+质粒中,并对608bp插入片段的顺序进行了双向测定。结果表明该克隆片段包含了小麦HMW谷蛋白亚基基因启动子CAAT box下游区域以及为谷蛋白肽链N-末端166个氨基酸编码的区域,通过与其它小麦中已知HMW谷蛋白亚基基因结构进行比较,表明本文扩增得到的顺序代表一个新的HMW谷蛋白亚基基因。The gene fragments of wheat (Triticum aestivum L. cv. Yang Mai No.4) high-molecular-weight (HMW) glutenin subunit were amplified in vitro using two chemically synthetic oligonucleotide primers and Taq DNA polymerase after 35 cycles of polymerase chain reaction. The amplified fragments were cloned into plasmid Bluescript SK^+ and the sequence of 608 bp inserted fragment was obtained in two directions.The sequence information has shown that the 608 bp fragment included downstream region of CAAT box and coding region for 166 N-terminal AA of wheat HMW glutenin subunit. As compared with HMW glutenin subunit genes from other species of wheat, it shows that the fragment amplified is a new HMW glutenin gene fragment.

关 键 词：聚合酶链反应 小麦 谷蛋白 

分 类 号：S512.101[农业科学—作物学]