“自身模板引物”PCR法构建蟋蟀神经肽Grb-AST_7基因串联体  

Cricket Grb-AST7 Concatemer Constructed by "Self Template-primer" PCR

作　　者：查笑君[1,2] 马伯军[1] 潘建伟[1] 黄俊生[2] 

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004 [2]中国热带农业科学院环境与植物保护研究所,海南儋州571737

出　　处：《安徽农业科学》2010年第13期6683-6685,共3页Journal of Anhui Agricultural Sciences

基　　金：国家科技支撑计划项目(2007BAD48B02);农业科技成果转化项目(2007GB23260410);中央级公益性科研院所基本科研业务费专项(2007HZS1J007)

摘　　要：[目的]通过改进的"自身模板引物"PCR法构建Grb-AST7基因串联体。[方法]根据蟋蟀神经肽Grb-AST7氨基酸序列设计合成2条引物,运用改进的"自身模板引物"PCR法进行拼接。[结果]通过拼接,获得了全长570bp、含17个拷贝的Grb-AST7基因串联体。拼接条件以拼接时间20min,25μlPCR体系中含2μl模板引物较合适。[结论]该研究为下一步进行Grb-AST7基因表达及其生物学活性分析奠定了基础。[Objective]Grb-AST7 concatemer including 17 copies of Grb-AST7 was constructed by ＂self template-primer＂ PCR. [Method]Two primers of which were synthesized based on the amino acid sequence of Gryllus bimaculatus’ Grb-AST7. [Result]Through splicing,a length of 570 bp,containing 17 copies of the Grb-AST7 gene repeats were obtained. Appropriate primer splicing conditions were splice time was 20 min,25 μl PCR system,containing 2 μl template. [Conclusion]The results laid a foundation for the future studies on Grb-AST7 gene expression and bio-activity analysis.

关 键 词：Grb-AST7 “自身模板引物”PCR 基因串联体 

分 类 号：Q969.263.4[生物学—昆虫学]