叶用莴苣热激蛋白90_LsHsp_省略_0_基因的克隆和其在热激下的表达_任月.pdfVIP

2013,46(16):3514-3522 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2013.16.023 102206 Molecular Cloning and Expression Analysis of Heat-Shock-Protein90 (LsHsp90) Gene from Leaf Lettuce (Lactuca sativa L.) Under Heat Shock REN Yue, HAN Ying-yan, LI Ting, HAO Jing-hong, FAN Shuang-xi (College of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206) Abstract: Objective LsHsp90 gene was cloned and used to analyze the expression profiles under heat shock stresses, in order to provide a foundation for investigating heat tolerant mechanisms of molecular regulation in stress tolerance of Lactuca sativa L.Method With homology cloning approaches coupling with RACE techniques, the full-length cDNA of LsHsp90 was cloned from total RNA of the leaves of L. sativa L. Real-time quantitative PCR (qRT-PCR) was used to analyze the expression patterns of this gene in leaves after heat shock unter 37 and 42 .ResultThe full cDNA sequence of LsHsp90 is 2 330 bpcontaining a 2 097 bp open reading flame (ORF) and encoding 698 amino acid residues with a predicted molecular mass of 79.8 kDThe LsHsp90 protein contains a predicted ATPase site and a Hsp90 conservative structure domain, which is highly conserved in plants and especially similar to Hsp90 from Ageratina adenophora (EU269070.1) and Arabidopsis thaliana (AY081302.1). Phylogenetic analysis showed that LsHsp90 and Hsp90 from Ageratina adenophora (EU269070.1) were clustered into one group. Real-time PCR analysis showed that, the expression of LsHsp90 in leaves of two varieties was upregulated under 37 . LsHsp90 was induced down regulation in heat-sensitive variety S106 under 42 ,