摘要
目的筛选具启动活性的新生隐球菌荚膜相关基因CAP10编码区上游5′侧翼序列(启动子序列),为进一步研究其调控机制打下基础。方法利用氯霉素乙酰基转移酶(CAT)的编码基因为报告基因,构建了含不同长度的新生隐球菌荚膜相关基因CAP10编码区上游5′侧翼序列和报告基因的重组体,转化酵母,ELISA方法检测各转化子的CAT表达活性,并通过比较找到CAP10启动序列。结果发现含CAP10编码区上游303bp、390bp、500bp、951bp的转化子具有明显的CAT表达活性,而含202bp、102bp的转化子无明显的CAT表达活性。结论CAP10上游5′侧翼端303bp～0bp是具完整启动活性的最小单位。CAP10上游5′侧翼端202bp～303bp内含CAP10启动所必需的序列。
Objective To find pronoter sequence of Cryptococcus neoformans capsule-associated gene CAP10, and so to provide a foundation for further experimental study on the mechanism of regulation. Methods Constructing recombinant plasmids that contain the different length of CAP10 coding region 5＇ upstream flanking sequence, following a gene that codes the chloramphenicol acetyltransferase（CAT） as a reporter gene. The recombinant plasmids were chemically transfected into yeast. The CAT activity of the reporter gene was assessed by ELISA to find G4P10 effective promoter region. Results The plasmids with -303 bp, -390 bp, -500 bp, -951bp length of CAP10 coding region 5＇ upstream flanking sequence all had significant CAT expressing activity. Those with -202 or -102 bp show no CAT expressing activity. Conclusion -303 bp - -0 bp length in CAP10 5＇ upstream flanking sequence acts the smallest intact promoter sequence. -202 bp- -303 bp length contains the necessary sequence for the CAP10 promoter activity.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第3期199-202,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金(30270080)资助
关键词
新生隐球菌
荚膜
基因
启动子
Cryptococcus neoformans
Capsule
Gene
Promoter
