Molecular Cloning and Sequence Analysis of the Toc33 cDNA in Brassica napus Line Cr3529  

作　　者：HUYuan-hui WANGMao-lin ZHANGNian-hui LIAOWen-tao ZHAOYun 

机构地区：[1]CollegeofLifeScience,SichuanUniversity,Chengdu610064,P.R.China

出　　处：《Agricultural Sciences in China》2005年第4期252-256,共5页中国农业科学（英文版）

基　　金：This work was supported by the National Natural Science Foundation of China(30170500);the Training Foundation ofthe Academic Leader ofSichuan Province,P.R.China.

摘　　要：Two primers, designed according to the Arabidoposis thaliana Toc33 sequence, were used to amplify the full coding region of the Toc33 cDNAs in leaves of a chlorophyll-reduced (Cr) mutant Cr3529 and its wild type 3529, Brassica napus, by RT-PCR technique. The RT-PCR results showed that the fragment homologous to Toc33 was expressed in Cr3529 as well as 3529 seedlings. PCR fragments were inserted into the pMD18-T vector and transferred into E. coli, then two cDNA clones, BnToc33-c and BnToc33, were obtained. Sequence analysis showed that the two sequences were 894 bp and the nucleotide and the deduced amino acid sequences were highly homologous to those of A. thaliana. There were three diverged nucleotides between the Cr3529 and the 3529 Toc33 cDNAs, i.e., GGT/AGT, TTG/TTT, AGG/AGT, all of which belonged to missense mutation. The amino acid replacement ((Leu/Phe) caused by TTG/TTT mutation located in the membrane anchor domain may result in chlorophyll-reduced character in Cr3529.Two primers, designed according to the Arabidoposis thaliana Toc33 sequence, were used to amplify the full coding region of the Toc33 cDNAs in leaves of a chlorophyll-reduced (Cr) mutant Cr3529 and its wild type 3529, Brassica napus, by RT-PCR technique. The RT-PCR results showed that the fragment homologous to Toc33 was expressed in Cr3529 as well as 3529 seedlings. PCR fragments were inserted into the pMD18-T vector and transferred into E. coli, then two cDNA clones, BnToc33-c and BnToc33, were obtained. Sequence analysis showed that the two sequences were 894 bp and the nucleotide and the deduced amino acid sequences were highly homologous to those of A. thaliana. There were three diverged nucleotides between the Cr3529 and the 3529 Toc33 cDNAs, i.e., GGT/AGT, TTG/TTT, AGG/AGT, all of which belonged to missense mutation. The amino acid replacement ((Leu/Phe) caused by TTG/TTT mutation located in the membrane anchor domain may result in chlorophyll-reduced character in Cr3529.

关 键 词：Brassica napus Protein translocation CHLOROPLAST MUTANT Sequence analysis 

分 类 号：S565.4[农业科学—作物学]