Engineering, cloning, and expression of genes encoding the multimeric luteinizing-hormone-releasing hormone linked to T cell determinants in Escherichia coli.
Two synthetic genes were designed and engineered to encode for multimeric luteinizing-hormone-releasing hormone (LHRH) peptides linked to T cell determinants. These genes were cloned into the prokaryotic expression vectors under control of strong inducible promoters, to overexpress the multimeric LHRH peptides as recombinant proteins. Multimeric LHRH-T cell peptides were expressed as insoluble inclusion bodies in Escherichia coli cultures. Cell extracts containing the recombinant proteins showed immunoreactivity on Western blots with monoclonal antibody recognizing the native hormonal peptide. These gene constructs have potential applications in therapy of sex-steroid-hormone-dependent cancers.
Gupta
Jagdish C
JC
The Talwar Research Foundation, New Delhi, India. jagdishgupta2@rediffmail.com
Raina
Komal
K
Talwar
G P
GP
Verma
Rashmi
R
Khanna
Navin
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D005813:Genes, Synthetic; D005818:Genetic Engineering; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics* / Q000378:metabolism; D002479:Inclusion Bodies / Q000737:chemistry; D008297:Male; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000737:chemistry / Q000502:physiology*
D010455:Peptides; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D007987:Gonadotropin-Releasing Hormone