木薯淀粉合成酶SSS Ⅱ基因片段的克隆与分析

摘要

[目的]建立木薯块根RNA的提取方法,并克隆和鉴定淀粉合成酶SSSⅡ基因核心片段.[方法]采用改良的CTAB法提取木薯块茎RNA,反转获得cDNA模板,同时基于已知植物的SSSⅡ基因同源性,设计简并引物,进行RT-PCR扩增,通过Blast在线检索比对对基因功能进行了初步鉴定.[结果]所获得片段即为木薯SSSⅡ基因的核心序列.[结论]该研究为木薯淀粉合成酶SSSⅡ基因全长cDNA序列的克隆及其反义载体的构建奠定了基础,同时为实现优质淀粉的代谢工程提供了理想的候选基因.%[ Objective] To establish an efficient method to extract RNA from cassava,and clone the core sequence of SSS II gene. [ Method] The cassava RNA was obtained using the improved CTAB method, which was then reverse transcripted into cDNA. Degenerate primers were designed based on the homology property of known SSS II sequences in other plant species. A fragment was amplified with the previously mentioned Cdna as template and the degenerate primers through Polymerase Chain Reaction (PCR). [Result] After online blasting in NCBI.the sequence was i-dentified to be the core fragment of cassava SSS II gene. [ Conclusion] The research would lay the original basis for the cloning of the cassava SSS II full length cDNA sequence and construction of its anti-sense vector,which could further provide proper candidate genes for the development of starch metabolic engineering.