天山雪莲rbcS基因启动子的克隆及序列分析  

Cloning and Sequence Analysis of Promoter Region of Saussurea involucrata Kar.et Kir rbcS Gene

作　　者：张亚敏[1] 杨晶[1] 王爱英[1] 祝建波[1] 

机构地区：[1]石河子大学生命科学学院,石河子大学农业生物技术重点实验室,新疆石河子832003

出　　处：《西北农业学报》2014年第5期146-152,共7页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金资助项目(31160049)

摘　　要：以天山雪莲(S.involucrata Kar.et Kir.)为材料,根据已获得的天山雪莲1,5-二磷酸核酮糖羧化/加氧酶小亚基基因组序列设计引物,采用热不对称交错PCR(TAIL-PCR)和高效TAIL-PCR(hiTAIL-PCR)扩增到rbcS基因5′上游启动子序列,长为1 668bp。用PlantCare和PLACE软件序列分析表明,该序列具有启动子的基本元件TATA-box、CA AT-box,包含多个胁迫诱导元件,如光诱导元件、赤霉素、低温诱导元件,昼夜节律调控元件等。构建pBI-PsikrbcS-GUS植物表达载体,并成功将其转化进入农杆菌。sikrbcS基因启动子的克隆与分析为进一步研究雪莲1,5-二磷酸核酮糖羧化/加氧酶小亚基基因的表达调控奠定了基础。Based on the known sequence of rbcSgene we designed nested specific primers,and the promoter sequence of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit gene fromSaussurea involucrata Kar.et Kir.genomic DNA was obtained by TAIL-PCR and hiTAIL-PCR.Nucleotide sequencing analysis showed that the full length of the fragment is 1 668bp.The functional elements were analyzed by PLACE and Plant Care programs.The promoter sequence contains the basic elements：TATA-box,CAAT-box and stress-induced elements,light responsive element,gibberellin responsive element,low-temperature responsiveness element and regulatory element involved in circadian control.The plant expression vector pBI-PsikrbcS-GUS was constructed,and then transformed into agrobacteria successfully.Cloning and analyzing of the promoter region of sikrbcSgene made substantial basis for further research on the mechanism of regulation and expression of Saussurea involucrata Kar.et Kir ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit gene.

关 键 词：天山雪莲 RBCS 热不对称交错PCR 高效TAIL-PCR 启动子 序列分析 

分 类 号：Q789[生物学—分子生物学]