Cloning and characterization of cotton heteromeric acetyl-CoA carboxylase genes  

作　　者：Qiao Zhixin Liu Jinyuan 

机构地区：[1]Laboratory of Molecular Biology and Protein Science Laboratory of the Ministry of Education, Department of Biological Sciences andBiotechnology, Tsinghua University, Beijing 100084, China

出　　处：《Progress in Natural Science:Materials International》2007年第12期1412-1418,共7页自然科学进展·国际材料（英文版）

基　　金：Supported by the State Key Basic Research and Development Plan of China (Grant No .2004CB117303);the Hi-Tech Research and DevelopmentProgramof China (2002AA207006);the National Natural Science Foundation of China (Grant Nos .30170080,39770078)

摘　　要：Heteromeric acetyl-coenzyme A （CoA） carboxylase （ACCase） catalyzes the formation of malonyl-CoA from acetyl-CoA. It plays an essential role in fatty acid synthesis in prokaryotes and most of plants. The heteromeric ACCase is composed of four subunits： biotin carboxylase （BC）, biotin carboxyl carrier protein （BCCP）, and α- and β-subunits of carboxyltransferase （α- and β-CT）. In this study, we cloned five novel genes encoding the subunits of heteromeric ACCase （one BC, BCCP and β-CT, and two a-CTs） from cotton （ Gossypiurn hirsuturn cv. zhongmian 35 ） by RACE-PCR. The deduced amino acid sequence of these cDNAs shares high similarity with other reported heteromeric ACCase subunits. The phylogenetic analysis indicated that the different subunits of heteromeric ACCase were grouped in a similar pattern. Southern blot analysis revealed the milti-copy patterns of these heteromeric ACCase genes in cotton genome. Semi-quantitative RT-PCR demonstrated that heteromeric ACCase genes were constitutively expressed in all of the cotton tissues, but the transcripts accumulated at a relatively low level in roots. To our knowledge, this is the first report about characterization of the heteromeric ACCase genes in cotton.Heteromeric acetyl-coenzyme A (CoA) carboxylase (ACCase) catalyzes the formation of malonyl-CoA from acetyl-CoA. It plays an essential role in fatty acid synthesis in prokaryotes and most of plants. The heteromeric ACCase is composed of four subunits: biotin carboxylase (BC), biotin carboxyl carrier protein (BCCP), and α- and β-subunits of carboxyltransferase (α- and β-CT). In this study, we cloned five novel genes encoding the subunits of heteromeric ACCase (one BC, BCCP and β-CT, and two α-CTs) from cotton (Gossypium hirsutum cv. zhongmian 35) by RACE-PCR. The deduced amino acid sequence of these cDNAs shares high similarity with other reported heteromeric ACCase subunits. The phylogenetic analysis indicated that the different subunits of heteromeric ACCase were grouped in a similar pattern. Southern blot analysis revealed the milti-copy patterns of these heteromeric ACCase genes in cotton genome. Semi-quantitative RT-PCR demonstrated that heteromeric ACCase genes were constitutively expressed in all of the cotton tissues, but the transcripts accumulated at a relatively low level in roots. To our knowledge, this is the first report about characterization of the heteromeric ACCase genes in cotton.

关 键 词：heteromeric ACCase COTTON gene cloning fatty acid synthesis. 

分 类 号：Q78[生物学—分子生物学]