摘要
在已知油茶乙酰COA酰基转移酶基因EST序列基础上,设计6条特异引物,以油茶优良无性系‘湘林1号’种仁总RNA为模板,通过反应体系和反应条件优化,最终扩增出一条700 bp左右的目的片段,将该片段回收、克隆、测序,获得718 bp的cDNA序列,将该序列与油茶乙酰COA酰基转移酶基因进行序列拼接,确定了油茶乙酰COA酰基转移酶基因的全长cDNA序列,将该序列提交至GeneBank,登录号为GU594059。油茶乙酰COA酰基转移酶基因全长cDNA序列为1 495 bp,含有一个1 227 bp的ORF,编码408个氨基酸残基。在氨基酸序列水平上,该基因与胡黄连(P.kurrooa)的乙酰COA酰基转移酶基因的相似性最高,为86%,与稻瘟病菌(M.grisea)的最低,为65%。通过在线预测,油茶乙酰COA酰基转移酶基因编码蛋白的等电点pI为6.19,分子量Mw为41 628.6 Da。本研究为揭示油茶油脂合成规律和油茶的分子育种提供了理论依据和科学基础。
Based on sequence information of acetyl-CoA C-acetyltransferase gene, six pairs of primers in conserved regions of this gene were designed. Total RNA of Xianglin No. 1 seeds was used as template, one fragment of 718 bp was successfully amplified by 5'RACE strategy. Then the full eDNA of acetyl-CoA C-aeetyltransferase gene was determined by assemblying the 5' sequence and known EST sequence that had been registered under GenBank (No. GU594059). The acetyl-CoA C-aeetyltransferase gene of Camellia oleifera was 1 495 bp long, including a completed ORE of 1 227 bp, encoding 408 amino acids. On the amino acid sequence level, this gene showed the highest similarity (86G) with acetyl-CoA C-acetyltransferase gene of P. kurrooa, and the lowest (65%) with M. grisea. In addition, the predicted pI and Mw of Camellia oleifera acetyl-CoA C-acetyltransferase gene was 6. 19, 41 628. 6 Da, respectively.
出处
《中南林业科技大学学报》
CAS
CSCD
北大核心
2011年第8期108-112,共5页
Journal of Central South University of Forestry & Technology
基金
国家"十一五"油茶科技支撑计划(2009BADB1B01
2009BADB1B02)
湖南省自然科学基金项目(10JJ4022)
中南林业科技大学青年基金项目(2009004A)
关键词
油茶
乙酰CoA酰基转移酶
快速扩增CDNA末端
生物信息学分析
Camellia oleifera
aeetyl-CoA C-acetyltransferase gene
rapid amplification of cDNA ends (RACE)
bioinformatics analysis