油桐异质型乙酰辅酶A羧化酶accA亚基全长cDNA克隆及序列分析

摘要

In order to reveal sequence features and structural function of accA gene in Vernicia fordii, and to lay a foundation for further research, development and utilization, taking nearly ripe seeds in V. fordii as materials, the speciifc primers were designed according to the results of transcriptome sequencing analysis of V. fordii, and full-length cDNA sequences of accA gene were cloned by RT-PCR technique. The results show that full-length cDNA sequence of accA gene is 2 313bp, encoding 770 amino acids. Iso electric point (pI) of accA protein is 8.48, the relative molecular mass is 85 902.7 Da, and the stability coefifcient is 37.33. The results of bioinformatics analysis show that, the protein has four obvious trans-membrane domains, and is an unstable non-secretory protein. The results of motif research show that a Carboxyl transferase domain and a ACCA functional domain are located inαsubunit. The tertiary structure of this protein has 14αhelixes, and appears a spherical body with a extended part. The gene has been submitted to the GeneBank and named by VfCTα.%为了揭示油桐乙酰辅酶A羧化酶accA基因的序列特点和结构功能，从而为该基因的深入研究和开发利用奠定基础，以葡萄桐的近成熟种子为材料，根据油桐转录组测序结果设计引物，采用RT-PCR技术克隆了油桐乙酰辅酶A羧化酶α亚基全长cDNA序列。测序结果表明：该基因cDNA序列全长2313 bp，编码770个氨基酸；以其推导出的α亚基氨基酸序列蛋白的等电点pI为8.48，相对分子量为85902.7 Da，稳定系数为37.33。生物信息学分析结果表明：此蛋白含有4个比较明显的跨膜区，是一个不稳定的非分泌蛋白。模体搜索结果表明：在α亚基上有羧基转移酶域和ACCA功能域；其三级结构中有14个α螺旋，整体呈球型，并有一个向外延伸的结构。该基因已登录到GenBank，命名为VfCTα。