【作者】 程汉良； 姬南京； 彭永兴； 申欣； 吴陈晨； 许建和； 董志国；

【Author】 CHENG Han-liang,JI Nan-jing,PENG Yong-xing,SHEN Xing,WU Chen-chen,XU Jian-he,DONG Zhi-guo (Jiangsu Key Laboratory of Marine Biotechnology,Huaihai Institute of Technology,Lianyungang 222005,China)

【机构】 淮海工学院江苏省海洋生物技术重点实验室；

【摘要】 提取草鱼肝脏总RNA,采用RT-PCR和RACE方法扩增草鱼乙酰辅酶A羧化酶(ACC)基因的核心序列、3’末端和5’末端序列,克隆测序,将各片段序列组装得到草鱼ACC基因全长cDNA序列,并对所得到的序列进行生物信息学分析。结果表明,草鱼ACC基因的全长cDNA序列共7433bp,开放阅读框共7071bp,共编码2357个氨基酸。推测前28个氨基酸构成一个信号肽,编码的蛋白是非分泌蛋白,蛋白分子量为265KDa,等电点为6.27,pH 7.0时带26.414个负电荷。该氨基酸序列与斑马鱼的相似性为96%,基于不同生物ACC氨基酸序列构建了系统发育树,发现ACC序列在脊椎动物中很保守,相似性相当高。更多还原

【Abstract】 Isolate the total RNA from liver of grass carp,we got the core sequence and two terminal sequences of the gene ACC with the method of RT-PCR and RACE.By means of assembling all kinds of fragments,we obtained the full-length cDNA of Acetyl-CoA Carboxylase from grass carp.Bioinformatics was applied in the analysis of the sequence,the results were as follows:The full-length nucleotide sequence which covers 7433bp has an open reading frame of 7071 nucleotides encoding 2357amino acids,including a putative signal peptide of 28 amino acids.The protein encoded is non-secreted protein,with a calculated molecular mass of 265KDa,isolectric point of 6.72,with -26.414 charge at pH 7.0.The deduced amino acids sequence has a homology of 96%with zebrafish.Furthermore,a phylogenetic tree based on the amino acid sequence of ACC was reconstructed.The tree’s topology illustrated that the Acetyl-CoA Carboxylase is conservative among vertebrate,The identity percent is rather high.更多还原