Real-time PCR Cloning and Expression of EhKCR1 from Eutrema halophilum

Abstract

Abstract: 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.

Key words: Eutrema halophilum, EmKCR1 In silico cloning, Bioinformatic analysis, Real-time PCR

Xu Xiaojing, An Huiling, Wei Baiyang, Zhou Yijun, Feng Jinchao . Real-time PCR Cloning and Expression of EhKCR1 from Eutrema halophilum[J]. Biotechnology Bulletin, 2013, 0(4): 55-62.

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