[Prokaryote expression and western analysis of BcpLH gene of Chinese cabbage].
BcpLH gene preferentially expressed in folding leaf of Chinese cabbage contains dsRNA-binding domains. The cDNA of BcpLH gene was cloned into a His-fusion expression vector pET-28a (+) and was induced to express in E. coli strain BL21 (DE3). Then, the specific protein was partially purified and the rabbit was immunized to prepare the anti-serum. Meanwhile BcpLH cDNA was cloned into the pMAL-c2 containing the solubizing partner, and then the soluble protein generated. It was demonstrated from Western dot assay that the BcpLH protein was specific. The BcpLH active protein and its anti-serum made it possible to study RNA-binding activity and regulation mechanism in plant development.
Yu
X H
XH
Shanghai Institute of Plant Physiology, Chinese Academy of Sciences, Shanghai 200032.
Zheng
Z R
ZR
Liu
P L
PL
He
Y K
YK
D015153:Blotting, Western; D001937:Brassica / Q000235:genetics*; D015870:Gene Expression; D011387:Prokaryotic Cells / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis*
D016601:RNA-Binding Proteins; D011994:Recombinant Proteins