title: [Cloning and expression of a novel metallothionein gene LbMT2 from Limonium bicolor].
abstract: The full length cDNA of a novel metallothionein (LbMT2) gene was cloned from a cDNA library of Limonium bicolor. The LbMT2 gene cloned is 518 bp in length, which includes a 64 bp of 5' untranslated region (UTR) and a 205 bp of 3' untranslated region. This gene has an open reading frame (ORF) of 249 bp in length, encoding a protein of 82 amino acid residues with the molecular mass of 8.1 kDa and theoretical pI of 4.71. The expression of LbMT2 gene in L. bicolor in response to CuSO4, CdCl2, NaCl, cold, and PEG was further investigated using real time quantitative PCR. In both leaf and root of L. bicolor, the expression of LbMT2 was induced by CuSO4, CdCl2, NaCl, and cold, but inhibited by PEG stress. LbMT2 gene was inserted into a prokaryotic expression vector (pGEX-4T-2) to produce the recombinant expression vector pGEX-LbMT2. The expression of LbMT2 in E. coli BL21 was induced with IPTG, which produced a protein band with expected size of 35 kDa on SDS-PAGE.
authors.affiliation: Northeast Forestry University, Harbin 150040, China. banqiaoying@yahoo.com.cn
authors.forename: Qiao-Ying
authors.initials: QY
authors.lastname: Ban
authors.forename: Gui-Feng
authors.initials: GF
authors.lastname: Liu
authors.forename: Yu-Cheng
authors.initials: YC
authors.lastname: Wang
authors.forename: Da-Wei
authors.initials: DW
authors.lastname: Zhang
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Jiang
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D019256:Cadmium Chloride / Q000494:pharmacology; D003001:Cloning, Molecular / Q000379:methods*; D019327:Copper Sulfate / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000235:genetics; D008668:Metallothionein / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D029606:Plumbaginaceae / Q000187:drug effects / Q000235:genetics*; D011092:Polyethylene Glycols / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012965:Sodium Chloride / Q000494:pharmacology
chemical_list: D011092:Polyethylene Glycols; D012965:Sodium Chloride; D008668:Metallothionein; D019256:Cadmium Chloride; D019327:Copper Sulfate