[Efficient expression of human C1-inhibitor in CHO cells by using a dicistronic expression vector].
The full-length cDNA fragment encoding human C1-INH was obtained by gene recombination techniques and a stable expression plasmid was constructed by inserting the human C1-INH cDNA into an efficient dicistronic expression vector pED. This plasmid was transfected into DHFR-deficient Chinese Hamster Ovary cells (CHO-dhfr-) by Lipofectin method, and the positive CHO-dhfr+ cell clones which stably express C1-INH was generated. Expression of C1-INH mRNA was detected by Northern blot. Expression of C1-INH protein was confirmed by immunoprecipitation and Western blot analysis. The level of expressed C1-INH was estimated at 0.4 microgram/ml by specific ELISA techniques. The biological activity of recombinant C1-INH was assayed by using the inhibition of C1 estrolytic activity.
Yang
H
H
Institute of Basic Medical Sciences, CAMS, Beijing.
Zheng
S
S
D000818:Animals; D016466:CHO Cells / Q000378:metabolism; D003174:Complement C1 Inactivator Proteins / Q000096:biosynthesis* / Q000235:genetics; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics; D014162:Transfection
D003174:Complement C1 Inactivator Proteins; D018076:DNA, Complementary; D012333:RNA, Messenger