Cloning and Sequence Analysis of a Osmotin-like Protein Gene from Atriplex canescens and Its Prokaryotic Expression

Abstract

Abstract: A osmotin-like protein gene was isolated based on the library from Atriplex canescens and its EST analysis, and named as AcOLP. The full length of AcOLP contained an open reading frame of 687 bp. It encoded a polypeptide of 229 amino acids, belonging to GH64- TLP-SF superfamily as a kind of PR-5 proteins. The AcOLP had 94% nucleotide sequence homology and 87% amino acid sequence homology to the sequence of osmotin-like protein from Atriplex centralasiatica, respectively. The accession number of AcOLP in GenBank was JN632587.1. Comparison of amino acid sequence with PR-5 proteins from various plants showed AcOLP possessed 16 cysteine residues that were conserved at their invariant and were presumably involved in disulfide bonding. Phylogenic analysis on the amino acid sequence of AcOLP with other plants showed that Atriplex canescens was closely related to Atriplex nummularia. AcOLP was inserted into the prokaryotic expression vector of pET-28a and expressed its fusion protein（about 29 kD）in Escherichia coli BL21（DE3）.

Key words: Atriplex canescens, Osmotin-like protein, Gene clone, Analysis of sequence, Prokaryotic expression

Wang Shengyang, Zhang Yong, Wang Jian, Liu Yanzhi, Liu Jinliang, Pan Hongyu . Cloning and Sequence Analysis of a Osmotin-like Protein Gene from Atriplex canescens and Its Prokaryotic Expression[J]. Biotechnology Bulletin, 2013, 0(2): 106-110.

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