[Cloning and expression of a novel metallothionein gene LbMT2 from Limonium bicolor].
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.
Ban
Qiao-Ying
QY
Northeast Forestry University, Harbin 150040, China. banqiaoying@yahoo.com.cn
Liu
Gui-Feng
GF
Wang
Yu-Cheng
YC
Zhang
Da-Wei
DW
Jiang
Li-Li
LL
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
D011092:Polyethylene Glycols; D012965:Sodium Chloride; D008668:Metallothionein; D019256:Cadmium Chloride; D019327:Copper Sulfate