关键词: 羊草;几丁质酶;RACE技术;生物信息学;原核表达

Abstract:

Based on the reported EST from Leymus chinensis, the chitinase ClassⅡgene (GenBank accession number GQ397277) from leaves of Leymus chinensis under 100 mmol/L Na2CO3 stress, named LcChi2 gene, was cloned by RACE. The sequencing and bioinformatic analysis revealed that the opening reading-frame (ORF) of this gene was 771 bp and encoding 256 amino acid. LcChi2 protein belonged to chitinase ClassⅡ, Family 19. The molecular weight of the protein was 27.4 kDa and the isoelectric point was 8.67. LcChi2 protein was highly identified with chitinase in Triticum aestivum, Hordeum vulgare, Secale cereale and Oryza sativa, sharing 96.5%、96.5%、95.2% and 80.5% homology, respectively, indicating that they had similar biological functions. The gene was expressed in E.coli for preparation of its polyclonal antibody used for detection of transgenic plant. The data predicted by bioinformatics are helpful for further analysis of LcChi2.

Key words: Leymus chinensis, chitinase, RACE, bioinformatics, prokaryotic expression