摘要
以山东生态型盐芥(EutremA halophilum)为试材,采用电子克隆及RT-PCR的方法,从中获得一个β-酮脂酰-CoA还原酶基因全长cDNA,命名为EhKCRI,GenBank登录号为JQ389855。EhKCR1cDNA长1 152 bp,含954 bp开放阅读框架,编码318个氨基酸。采用生物信息学的手段对EhKCR1蛋白的保守结构域、疏水性和二级结构等进行了分析。EhKCR1蛋白具有NADH结合结构域[G(X)3GXG(X)3A(X)3A(X)2G]负责裂解的关键结构域[Y(X)3K]。序列比对和系统进化分析表明,盐芥EhKCRl基因与来源于拟南芥和油菜的KCR1亲缘关系最近,与单子叶植物来源的KCR1亲缘关系较远。Real-time PCR分析表明EhKCRI受ABA和干旱明显诱导。推测EhKCRI与盐芥的抗逆有关。
A cDNA was cloned from Eutrema halophilum （ Shangdong ecotype ） using in silico cloning and RT-PCR methods. This gene was predict coding β-ketoacyl-CoA reductase, named as EhKCR1, with GenBank accession number JQ389855. The full length is 1 152 bp and contains a complete ORF with 954 bp encoding 318 amino acid. The conservative domain, hydrophobicity and second structure of EhKC1 protein were predicted by bioinformatic analysis. EhKCR1 has the putative NADH binding motif [ G （ X ） 3GXG （ X ） 3A （ X ） 3A （ X ） 2G ] and the essential catalytic motif （ Y （ X ） 3K ） . Alignment of predicted amino acid sequence of EhKCR1 with other plants and phylogenetic analysis of various EhKCR1 homologues found that EhKCR1 was very close to that from Arabidopsis thaliana and Brassica napus, but far from to EhKCR1 of monocotyledon. The results of Real-time PCR indicated that EhKCR1 was induced by ABA and drought. Therefore EhKCR1 may be connected with the strong stress tolerence of Eutrema halophilum.
出处
《生物技术通报》
CAS
CSCD
北大核心
2013年第4期55-62,共8页
Biotechnology Bulletin
基金
国家自然科学基金项目(31100287)
中央高校基本科研业务费专项资金(1112KYZY43)
中央民族大学"985"项目(MUC98504-14
MUC98507-08)
