Cloning, Expression,
Purification and Activity of the hsBLyS
LIU ping, ZHANG Shuang-Quan*, YAN Xiao-Mei, HUANG
Hao-Jie, ZHOU Kai-Ya
( Academy of Life Science, Nanjing Normal University, Nanjing 210097,
China )
Abstract The full
length cDNA of human B lymphocyte stimulator (hBLyS) was amplified by using PCR
method from cDNA library of human placenta. After purifying and sequencing, the
DNA fragment of functional domain of hBLyS (hsDNA fragment)was
amplified by using nested PCR method from the PCR product. The prokaryotic
expression plasmid pET-30a(＋)/hsBLyS was constructed with
recombinant DNA techniques after purifying and identifying the hsDNA fragment.
Then the plasmid pET-30a(＋)/hsBLyS was transformed into
λDE3 cells and the recombination protein was found to be highly expressed； the expression product was
purified by affinity chromatography gel, Ni2＋-IDA, made in our laboratory.
The experimental results showed that the sequence of the PCR product was
identical with the published hBLyS cDNA sequence and purity of the
recombination protein we obtained was high. The activity of the purified
recombination protein was very significant in the proliferation test of B
lymphocytes.
Key words hsBLyS； cell apoptosis； transmembrane protein； fusion protein； cell proliferation
*Corresponding author： Tel, 86-25-3598216； e-mail, [email protected]