Cloning and Expression of Lipoxygenase Homology Protein from Leymus chinensis

doi:10.7668/hbnxb.2015.04.007

Abstract

Abstract: In order to know the relationships between Lipoxygenase and salt stress,a full length cDNA named LcLHP related to saline-alkali tolerance was cloned from Leymus chinensis by RT-PCR and RACE(Rapid amplification of cDNA ends). LcLHP gene (GenBank:KJ472894) with 537 bp open reading frame encoded a protein with a predicted length of 178 amino acids.LcLHP has PLAT/LH2 structural domain,which is induced by stress.Homology analysis showed that the deduced amino acid sequence of LcLHP shared high homology with PLAT/LH2 protein family of Spartina alterniflora and PLAT stress protein.Protein function prediction showed N terminal of LcLHP may have signal peptide,act as enzyme.The expression of LcLHP was analyzed by RT-PCR under salinity-alkalinity stressed condition.With the extension of the salinity-alkalinity stressed time,the expression of LcLHP increased from 0 to 12 h in the leaf,and then decreased,the peak of expression level arrived at 12 h.Construction of pGEX-LcLHP expression vector,IPTG induced prokaryotic expression,we got GST-LcLHP fusion protein and verified by Western Blot.

Key words: Leymus chinensis, Saline-alkali stress, LcLHP, RACE cloning, Expression analysis

CLC Number:

ZHANG Yang, SUN Tian-xu, GUO Bo-yu, TANG Ming-wei, LI Yu-hua, XIE Li-nan. Cloning and Expression of Lipoxygenase Homology Protein from Leymus chinensis[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(4): 35-42. doi: 10.7668/hbnxb.2015.04.007.

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