Construction, purification and characterization of a recombinant baculovirus containing the gene for alpha subunit of human chorionic gonadotropin.
A cDNA encoding the alpha subunit of human chorionic gonadotropin, a placental glycoprotein hormone, was cloned downstream to the viral polyhedrin gene promoter of Autographa california nuclear polyhedrosis virus and the recombinant transfer vector was used to co-transfect Spodoptera frugiperda cells growing in culture. Recombinant baculovirus carrying the alpha hCG gene was detected and isolated after dot hybridization using supernatant from co-transfected cells. Recombinant vAc alpha hCG having a replacement of the viral polyhedrin gene, which is hyper-transcribed very late in the infection cycle, with the alpha hCG cDNA was purified after a single round of plaque purification. Insect cell culture infected with vAc alpha hCG, secreted high levels of hCG which was biologically active.
Nakhai
B
B
National Institute of Immunology, Shahid Jeet Singh Marg, New Delhi, India.
Sridhar
P
P
Talwar
G P
GP
Hasnain
S E
SE
D016367:Baculoviridae / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005822:Genetic Vectors; D015292:Glycoprotein Hormones, alpha Subunit / Q000235:genetics*; D006801:Humans; D015183:Restriction Mapping
D015292:Glycoprotein Hormones, alpha Subunit; D004247:DNA