生命科学研究2011，Vol.15Issue(6)：485-492,8.
云南红梨PyMT基因的克隆、原核表达和功能分析
Cloning, Prokaryotic Expression and Function Analysis of PyMT Gene from Yunnan Red Pear (Pyrus pyrifolia)
摘要
Abstract
Metallothionein is a kind of ubiquitous and multifunctional protein in organism. PyMT (Pyrus pyrifolia metallothionein) genomic sequences were amplified according to the EST sequence of the red pear ' Yunhong-1' s light-specific supression substration hybridization library from four cultivars of Yunan red pear and these sequences were analyzed. The PyMT cDNA sequence of 'Yunhong-1' was amplified by RT-PCR and was designated PyMTl (Pyrus pyrifolia metallothionein 1). Prokaryotic expression vector pGEX-4T-1-PyMT1 was constructed and was transformed into E. Coli BL21 for expression. The growth curves of E. Coli. Transformed with pGEX-4T-l-PyMR1 and the control were obtained by growing them in liquid medium with different concentration of various metal ions. Sequence analysis showed that PyMTl belongs to the typical Metallothion 2. Results of phylogenic analysis showed that PyMTl was at the same evolutionary branch with MdMT. The SDS-PAGE results display that the expressed proteins are consistent with the anticipated size. Results of the growth curve show that PyMTl has a certain tolerance to heavy mental ions such as Zn, Cu and Cd in vivo. The results will provide a foundation for further purification, structure and function research pf PyMTl.关键词
云南红梨/PyMT/序列分析/原核表达/重金属抗性Key words
Yunnan red pear/PyMT/sequence analysis/prokaryotic expression/heavy mental resistance分类
生物科学引用本文复制引用
樊磊,张晓东,陈丽梅,苏俊,舒群,李昆志..云南红梨PyMT基因的克隆、原核表达和功能分析[J].生命科学研究,2011,15(6):485-492,8.基金项目
国家自然科学基金资助项目(31060258) （31060258）
