甘蓝型油菜雄性不育基因BnaX.MS1.a的克隆及其amiRNA载体的构建

›› 2010, Vol. 22 ›› Issue (5): 552-557.

甘蓝型油菜雄性不育基因BnaX.MS1.a的克隆及其amiRNA载体的构建

刘振林^1,2,柯丽萍²,何海燕²,陈锦清^2,*

¹浙江师范大学化学与生命科学学院, 浙江金华321004；²浙江省农业科学院病毒学与生物技术研究所,浙江杭州310021

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-09-25 发布日期:2010-09-25

Molecular cloning of male sterile gene BnaX.MS1.a in Brassica napus and construction of a vector with MS1 amiRNA

LIU Zhen-lin;KE Li-ping;He Hai-yan;CHEN Jin-qing;*

¹College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China；²Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:1900-01-01 Revised:1900-01-01 Online:2010-09-25 Published:2010-09-25

摘要/Abstract

摘要： 根据拟南芥MS1基因已知保守序列设计特异性引物,分离克隆甘蓝型油菜中与拟南芥MS1基因同源的基因片段。应用PCR-Walking技术获得甘蓝型油菜核雄性不育基因BnaX.MS1.a,其全长3 424 bp。BnaX.MS1.a基因与拟南芥MS1基因全长序列同源性为60.3%,编码序列同源性为89.5%。根据基因序列比对结果推测开放阅读框长2 004 bp,编码667个氨基酸,分子量为765 kD,等电点为8.01。BnaX.MS1.a基因编码的氨基酸序列与已知拟南芥MS1基因编码的氨基酸序列同源性达93%。构建BnaX.MS1.a基因的amiRNA载体转化甘蓝型油菜,旨在研发核不育基因工程油菜。

关键词: MS1, BnaX.MS1.a, 甘蓝型油菜, amiRNA

Abstract: MS1 gene is a tapetum-specific expressed gene related with male sterility in Arabidopsis thaliana. According to the coding sequences of Arabidopsis MS1 gene, a series of primers were designed to clone the homologous sequences in Brassica napus. PCR-Walking method was used to clone the full sequence of BnaX.MS1.a gene of Brassica napus. The total length of BnaX.MS1.a gene cloned here was 3 424 bp. The MS1 from Brassica napus shared a similarity of 89.5% in encoding sequence and 60.3% in full sequence with that from Arabidopsis thaliana. According to the results, it was deduced that it had an open reading frame (ORF) of 2 004 bp, encoding a protein of 667 amino acid, with protein molecular weight of 26 kD and isoelectric point of 8.01. The similarity between MS1 gene from Brassica napus and that from Arabidopsis thaliana was 93% in amino acid sequence. A plant expressed vector with the artificial miRNA was constructed for further studies on male sterility of Brassica napus.

Key words: MS1, BnaX.MS1.a, Brassica napus, amiRNA

刘振林;柯丽萍;何海燕;陈锦清;*. 甘蓝型油菜雄性不育基因BnaX.MS1.a的克隆及其amiRNA载体的构建[J]. , 2010, 22(5): 552-557.

LIU Zhen-lin;KE Li-ping;He Hai-yan;CHEN Jin-qing;*. Molecular cloning of male sterile gene BnaX.MS1.a in Brassica napus and construction of a vector with MS1 amiRNA[J]. , 2010, 22(5): 552-557.

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