石栗accD基因全长cDNA的克隆及序列分析

Cloning and analysis of full length cDNA sequence of accD gene from Aleurites moluccana

【目的】克隆石栗幼嫩叶片accD基因的全长cDNA序列,为今后利用该基因和提高油料作物种子含油量提供参考。【方法】根据已知植物麻疯树、蓖麻等accD基因的保守区域设计简并引物,以石栗幼嫩叶片为材料,利用简并PCR和RACE技术克隆accD基因的全长cDNA序列,扩增其编码序列。【结果】扩增获得的石栗accD基因全长cDNA序列长1789 bp,包含1509 bp的开放阅读框,可编码503个氨基酸,GenBank登录号为KC255250,并命名为amaccD。石栗accD基因序列经BLASTx比对后,发现其与巴西橡胶树、蓖麻和麻疯树氨基酸序列的同源性分别为90%、90%和89%。【结论】克隆获得的石栗accD基因全长cDNA序列与巴西橡胶树、蓖麻和麻疯树等具有较高同源性。

[Objective]The full length cDNA sequence of accD gene of acetyl coenzyme A carboxylase （ACCase） [3-CT subunit from Aleurites moluccana tender leaves was cloned to provide references for using this kind of gene and improving the oil content of oil-bearing crops seeds. [Method]Degenerate primers were designed based on the conserved region of known accD gene of plants, such as Jatropha curcas and Ricinus communis. Tender leaves of Aleurites moluccana were used as the materials. PCR and RACE technology were used to clone and amplify the full length cDNA sequence of accD gene. [Result]We obtained a 1789 bp sequence of full length cDNA of accD gene, which contained 1509 bp open reading frame and could encode 503 amino acids. The accession number of GenBank was KC255250, which was named as amac- cD. The results of BLASTx comparison showed that the homlolgy of the amino acid sequence among accD gene of Aleurites moluccana and the known homologous plants Hevea brasiliensis, Ricinus communis and Jatropha cureas was 90%, 90% and 89%, respectively. [Conclusion]The full length sequence of accD gene in Aleurites moluccana had high homology with Hevea brasiliensis, Ricinus communis and Jatropha eurcas.