Chicken Calcitonin Gene Recombinant Plasmid And Its Expression In Laying Hens In Vivo Study

Calcitonin is the main hormone in adjusting the metabolism of calcium to human or animals. Hence it is widely used to treat osteoporosis,,hypercalcemia and various bone disease,such as the Paget's disease.Most calcitonin in clinic use is extrated form salmon ultimobranchial gland or pig thyroid gland,so it is expensivejt had been produced though prokaryotic expression system,,but the deficiency in modification after translation restricted its application,In the research we attempted to construct two eukaryotic expression plasmids of chicken Calcitonin ,through which to study the possible mechanism of calcium metabolism ,and to explore a new solution to calcium deficiency by gene vaccine.A pair of PCR primers was designed according to the published chicken calcitonin sequence. Through RT-PCR, cDNA of the gene was got and then cloned into teukaryotic expression vector, pCEP4 and pcDNA3. Thus, two recombinant plasmids were injected into chickens. And a boost injection two weeks later. Blood samples were collected every two weeks. Serum concentration of CCT,PTH, E2, Ca and P were detected to study the expressing of pCEP4-cCT and pcDNA3-cCT.The results showed.1. The gene segment we got had only one nucleotide difference with the published chicken CCT gene.2. Two teukaryotic expression plasmid pCEP4-cCT and pcDNA3-cCT were success constructed.3. Compared with the controlled group both injection of pCEP4-cCT and pcDNA3-cCT caused the decline of serum Ca. But CT, PTH, E2 and P ascended. The results showed the plasnid had a significant effect.4. Compared with pcDNA3-CCT, the impressing of pCEP4-CCT maintained a longed time and better effect.