[Cloning and optimizing prokaryotic induced expression conditions of prolactin in White Goose].
The mature segment gene of prolactin (PRL) in White Goose was amplified from pituitary by RT-PCR and then cloned into the pMD18-T vector. Sequencing analysis showed that the cDNA has a length of 690 bp including the termination codon and encodes a protein composed of 230 amino acids, which differs from the published PRL cDNA sequence. There is a homology of 99.57% in base and 99.56% in amino acids with that of Wanxi White Goose, respectively. A prokaryotic expression vector, pET-32a(+), was used to construct the recombinant plasmid pET-32a(+)-PRL to produce protein. Having been induced by IPTGthe host cell carrying the recombinant plasmid expressed the recombinant PRL. The optimal condition for expression is 1 mmol/L IPTG at 37. Based on this condition, the expression rose to the highest level by 4 h of induction, accounting for 28.96% of the total bacterial protein.
Guo
Li
L
College of Animal Science and Veterinary Medicine, Heilongjiang August First Land Reclamation University, Daqing 163319, China. guoli_1213@163.com
Yang
Huan-Min
HM
Li
Peng
P
Kang
Bo
B
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics*; D005777:Geese / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D002479:Inclusion Bodies; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008969:Molecular Sequence Data; D011388:Prolactin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013696:Temperature
D007544:Isopropyl Thiogalactoside; D012313:RNA; D011388:Prolactin; D004262:DNA Restriction Enzymes