Cloning and Expression Analysis of CkP5CS Gene in Caragana korshinskii

doi:10.13560/j.cnki.biotech.bull.1985.2018-0349

Abstract

Abstract: The study aims to clone the full-length cDNA sequence of gene P5CS from Caragana korshinskii,analyze its tissue expression specificity and expression under the stresses of low temperature,high salt and PEG,and clarify its biological functions. The full-length cDNA of P5CS from C. korshinskii was isolated by RACE technique and analyzed by bioinformatics. Phylogenetic tree was constructed for studying homology with similar sequences. RT-PCR method was used to analyze the tissue expression specificity and the expression of CkP5CS under low temperature,high salt and osmotic stress. As results,the full-length cDNA sequence of P5CSgene of C. korshinskii was cloned and named as CkP5CS with a length of 2 604 bp,containing a 5'-UTR of 102 bp,a 3'-UTR of 363 bp,and a 2 139 bp opening reading frame,and it encoded 712 amino acids with the molecular weight of 46.8kD and theoretical isoelectric point of 8.6.The main secondary of the protein was a helix,extended strand and random coils. Comparison of amino acid sequences and phylogenetic analysis the CkP5CS showed high identity with those from Sophora davidii SdP5CS（85.1%）,Glycine max GmP5CS（84.2%）and Vigna unguiculata VuP5CS（82.6%）. Real-time PCR analysis revealed that CkP5CS expressed in roots,stems,leaves,indicating that there was almost no tissue specificity. The transcript level of CkP5CS was induced in response to low temperature,high salt and osmotic stress. As conclusion,CkP5CS gene isolated from C. korshinskii plays an important role in the adaptive process under low temperature,high salt and osmotic stress.

Key words: Caragana korshinskii, CkP5CS gene, molecular cloning, expression analysis

ZHANG Teng-guo, SHI Zhong-fei, KOU Ming-gang, ZHENG Sheng, WANG Juan. Cloning and Expression Analysis of CkP5CS Gene in Caragana korshinskii[J]. Biotechnology Bulletin, 2018, 34(9): 209-218.

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