胡麻异质型ACCase亚基基因的克隆与表达分析

为研究胡麻异质型乙酰辅酶A羧化酶BCCP、BC、α-CT和β-CT四个亚基基因的生物学功能,采用基因克隆、生物信息学和RT-PCR技术分别对胡麻accA、accB、accC和accD 4个基因进行分析。结果发现陇亚10号accA基因编码α-CT亚基,CDS序列全长为2364 bp,编码787个氨基酸;accB基因编码BCCP亚基,CDS序列全长为1173 bp,编码275个氨基酸;accC基因编码BC亚基,CDS序列全长为1574 bp,编码527个氨基酸;accD基因编码β-CT亚基,CDS序列全长为1137 bp,编码378个氨基酸,且4个基因编码的蛋白都是脂肪酸系数较高的亲水性蛋白。RT-PCR结果表明,胡麻accA、accB、accC和accD 4个基因在3个胡麻品种(高含油量的张亚2号、中含油量的陇亚10号和低含油量的R2-17)的不同时期不同组织的表达模式不同,accA、accB、accC和accD四个基因在R2-17、陇亚10号和张亚2号3个胡麻品种所有时期的不同组织中均有表达,但在种子中的表达量均明显高于其他组织,尤其在种子发育10~25 d油脂快速积累时期,4个基因的表达量都迅速增加并达到最高峰。研究表明异质型乙酰辅酶A羧化酶基因可能调控胡麻种子发育前期油脂的合成积累。

Molecular cloning and expression of heteromeric ACCase subunit genes from flax

In order to study the biological function of flax heterogeneous acetyl-CoA carboxylase BCCP, BC, α-CT and β-CT genes, we used the gene cloning technology RT-PCR and bioinformatics analysis, to analyze four genes of flax accA, accB, accC and accD. It was found that the accA gene of Longya No. 10 encodes the α-CT subunit, and the CDS sequence is 2364 bp in length and encodes 787 amino acids; the accB gene encodes the BCCP subunit, and the CDS sequence is 1173 bp in length and encodes 275 amino acids; the accC gene encodes the BC subunit, and the CDS sequence is 1574 bp in length and encodes 527 amino acids; the accD gene encodes a β-CT subunit, and the CDS sequence is 1137 bp in length and encodes 378 amino acids. The proteins encoded by these four genes are all hydrophilic proteins with high fatty acid coefficients. RT-PCR results show that in different tissues of the three flax varieties, Zhangya No.2 with high oil content, Longya No.10 with medium oil content, and R 2-17 with low oil content, the four genes of flax accA, accB, accC, accD have different expression modes. During the rapid oil accumulation period of seed development (10-25 d), the expression levels of these four genes increased rapidly and reached the highest peak. Studies have shown that the heterogeneous acetyl-CoA carboxylase genes may regulate the accumulation of lipid in flax seeds during early development.