Cloning, expression and identification of two glutathione S-transferase isoenzymes from Perna viridis

• Main Authors: Li, Zhenzhen, Chen, Rong, Zuo, Zhenghong, Mo, Zhengping, Yu, Ang, 陈荣, 左正宏, 郁昂
• Format: Article in Journal/Newspaper
• Language: English
• Published: ELSEVIER SCIENCE INC 2013
• Subjects: OYSTER CRASSOSTREA-GIGAS; MOLECULAR-CLONING; PI-CLASS; OMEGA-CLASS; MU-CLASS; MYTILUS-GALLOPROVINCIALIS; VENERUPIS-PHILIPPINARUM; GENE-EXPRESSION; CDNA CLONING; GST GENES; Crassostrea gigas
• Online Access: http://dspace.xmu.edu.cn/handle/2288/87998

Natural Science Foundation of Fujian Province [2008J0138]; program for Changjiang Scholars and Innovative Research Team in University [IRT0941] Glutathione S-transferases (GSTs; EC 2.5.1.18) are phase II enzymes involved in major detoxification reactions of xenobiotic in many organisms. In the present study, two classes of GSTs (PvGST1 and PvGST2) were cloned from P. viridis by rapid amplification of cDNA ends method. Sequence alignments and phylogenetic analysis together supported that PvGST1 and PvGST2 belonged to the pi and omega classes, respectively. The PvGST1 cDNA was 1214 nucleotides (nt) in length and contained a 618 nt open reading frame (ORF) encoding 206 amino acid residues, and had 46 nt of 5'-untranslated region (UTR) and a 3' UTR of 550 nt including a tailing signal (AATAAA) and a poly (A) tail. The molecular mass of the predicted PvGST1 was 23.815 kDa, with the calculated isoelectric point being 5.39. PvGST2 was 1093 bp, consisting of a 5' UTR of 13 bp, a 3' UTR of 246 bp and an ORF of 834 bp. The deduced protein was composed of 278 amino acids, with an estimated molecular mass of 32.476 kDa and isoelectric point of 8.88. Tissue distribution analysis of the PvGST1 and PvGST2 mRNA revealed that the GST expression level was higher in digestive gland and gonad, while lower in gill and mantle in both genders. Molecular modeling analysis of two GSTs implicated their various functions account for their different enzymatic features. (c) 2013 Elsevier Inc. All rights reserved.