四川大学学报(自然科学版)
四川大學學報(自然科學版)
사천대학학보(자연과학판)
JOURNAL OF SICHUAN UNIVERSITY
2000年
3期
446-450
,共5页
甘薯储藏蛋白%基因表达%大肠杆菌%SDS-PAGE
甘藷儲藏蛋白%基因錶達%大腸桿菌%SDS-PAGE
감서저장단백%기인표체%대장간균%SDS-PAGE
sporamin A%gene expression%Escherichia coli%SDS-PAGE
利用基因重组技术,将PCR扩增获得的甘薯储藏蛋白A基因的成熟肽编码区序列,插入到高效表达载体pTrcHisB中,构建成重组表达质粒pTHSPO-A.用限制酶切和PCR分析均表明重组质粒与预期结果相符.用IPTG诱导重组质粒转化的大肠杆菌TOP10,菌体总蛋白经12%SDS-PAGE分析,可观察到一个大小为10kD的蛋白质带,其分子量比预期值小.
利用基因重組技術,將PCR擴增穫得的甘藷儲藏蛋白A基因的成熟肽編碼區序列,插入到高效錶達載體pTrcHisB中,構建成重組錶達質粒pTHSPO-A.用限製酶切和PCR分析均錶明重組質粒與預期結果相符.用IPTG誘導重組質粒轉化的大腸桿菌TOP10,菌體總蛋白經12%SDS-PAGE分析,可觀察到一箇大小為10kD的蛋白質帶,其分子量比預期值小.
이용기인중조기술,장PCR확증획득적감서저장단백A기인적성숙태편마구서렬,삽입도고효표체재체pTrcHisB중,구건성중조표체질립pTHSPO-A.용한제매절화PCR분석균표명중조질립여예기결과상부.용IPTG유도중조질립전화적대장간균TOP10,균체총단백경12%SDS-PAGE분석,가관찰도일개대소위10kD적단백질대,기분자량비예기치소.
The coding sequence of mature sporamin A of sweet patato was amplffied by PCR and inserted into the expression vector pTrcHisB , resulting in the recombinant plasmid pTHSPO-A, which was confirmed by restriction and PCR analysis . The pTHSPO-A was transformed into Escherichia coli TOP10 . After induction with IPTG ,an additional protein band with MW of 10 kD was observed by SDS-PAGE.