水稻热激蛋白转录因子基因HsfA1aRNAi载体的构建
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水稻热激蛋白转录因子基因HsfA1aRNAi载体的构建(9000字)
摘 要：从水稻HsfA1a (heat shock transcription factor A1a)基因cDNA3'编码区扩增约500bp的RNAi顺式和反式片段，再从质粒pBI121中扩增出部分GUS片段用作RNAi的顺反片段之间的中间序列，然后依次连接到中间载体pBSK上，最后将整个RNAi片段插入到表达载体p1301M中，构建一个以CaMV35S启动子驱动表达的水稻HsfA1a基因RNAi表达载体，将其通过冻融法转入根瘤农杆菌EHA105，用于水稻的遗传转化，为鉴定水稻HsfA1a基因的功能奠定基础。
关键词：水稻；热激蛋白转录因子基因HsfA1a；RNAi载体；双酶切；PCR
Construction of RNAi expression vector for HsfA1a gene of rice
Abstract: To construct RNAi expression Vectors of the Rice HsfA1a gene, the sense and antisense fragments were amplified by PCR from the 3’ end of about 500bp coding sequences of cDNA clones in rice and ligated to both sides of partial GUS gene that was amplified from pBI121. At first, RNAi fragments were linked into vector pBSK, then cut down and linked into expression vector p1301M driven by CaMV35S promoter. The successfully constructed vector for HsfA1a gene was transformed into EHA105 by freeze-thawing method, which used for genetic transformation of rice. This work was a good start for further study on the functions of rice HsfA1a gene.
Key words: Oryza sativa; Heat shock transcription factor gene HsfA1aRNA; interference vector; Double digestion; Polymerase chain reaction
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