杜氏盐藻FLA8基因全长的克隆及鉴定

Cloning and identification of the full-length cDNA sequence of FLA8 gene from Dunaliella salina

目的:获得FLA8基因cDNA序列。方法:根据衣藻、小球藻等驱动蛋白-2亚基的氨基酸高度保守序列GYNGTIF、NEDPKDA设计一对简并引物,采用RT-PCR及3’RACE和5’RACE的方法扩增杜氏盐藻FLA8基因的全长。结果:克隆得到的杜氏盐藻FLA8基因cDNA全长序列为2612bp,序列分析显示其包括90bp的5’UTR、167bp的3’UTR以及2355bp的开放阅读框,其编码784个氨基酸残基,蛋白质的理论等电点为6.65,相对分子质量为85097。氨基酸序列同源性分析显示其与其他物种有较高的同源性:团藻(98%)、衣藻(99%)、小球藻(92%)。结论:得到杜氏盐藻驱动蛋白-2亚基FLA8的基因全长。

Aim：To get the full-length cDNA sequence of FLA8 gene from Dunaliella salina（D.salina）.Methods：A pair of degenerated primers was designed according to conserved homologous amino acid sequences of GYNGTIF and NEDPKDA of kinesin-2 motor subunit of FLA8 from microalgae and other organisms.The full-length cDNA of FLA8 gene was obtained by RT-PCR,3＇and 5＇RACE technology.Results：The results showed that the full-length cDNA sequence of the cloned FLA8 gene was 2 612 bp,possessing a 90 bp 5＇UTR,a 167 bp 3＇UTR and a 2 355 bp open reading frame encoding 784 amino acids.The theoretical pI value of FLA8 was 6.65 and the relative molecular weight was 85 097.Amino acid sequence homologous analysis indicated that FLA8 of D.salina shared high homology with other organisms,such as Volvox carteri f.nagariensis（98%）,Chlamydomonas reinhardtii（99%）,Micromonas（92%）.Conclusion：The full-length cDNA sequence of FLA8 gene from D.salina has been cloned.