酵母K.cicerisporus基因组中有启动子功能的DNA片段的克隆  

Cloning of DNA Fragments Possessing Promoter Function from Kluyveromyces cicerisporus

作　　者：徐国洲[1] 潘锋[1] 霍克克[1] 李育阳[1] 

机构地区：[1]复旦大学遗传学研究所遗传工程国家重点室验室

出　　处：《生物化学与生物物理学报》1998年第1期35-40,共6页

基　　金：国家高技术"863"计划

摘　　要：利用以３′－氨基糖苷磷酸转移酶基因（ＡＰＨＩ）为报告基因的一套启动子探针质粒ｐＳＫ－ｋａｎ４０１、ｐＳＫ－ｋａｎ１１０５、ｐＳＫ－ｋａｎ１２３８，从酵母Ｋｌｕｙｖｅｒｏｍｙｃｅｓｃｉｃｅｒｉｓｐｏｒｕｓ基因组中克隆到８个有较强启动功能的ＤＮＡ片段，分析出３′端ＤＮＡ序列，并在酵母Ｋｌｕｙｖｅｒｏｍｙｃｅｓｌａｃｔｉｓ中通过检测报告基因与３－磷酸甘油醛脱氢酶基因（ＧＡＰＤＨ）的ｍＲＮＡ表达量的比值，确定它们的功能强度，其中ｐＳＫ－ｋａｎ４０１－４１和ｐＳＫ－ｋａｎ１１０５－５１两个克隆的插入片段具有较强的启动子功能，并证明这两个片段在酵母Ｋ．ｃｉｃｅｒｉｓｐｏｒｕｓ中也有启动子功能。Using three yeast promoterprobe plasmids pSKkan401、pSKkan1105、pSKkan1238 with different reading frames, eight DNA fragments possessing high efficient promoter function have been cloned from K.cicerisporus. Their 3′DNA sequences have been analysed. The relative strength of their promoter function was studied by comparing their ability to promote the expression of the reporter gene APHI with that of glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) promoter in K.lactis. The results showed that the insertion fragments in pSKkan40141 and pSKkan110551 have stronger promoter function and they are also functional in K.cicerisporus Y179.

关 键 词：启动子 克隆 酵母 基因组 DNA片段 

分 类 号：Q782[生物学—分子生物学]