닭 정자를 벡터로 이용한 형질전환 닭 생산에 관한 연구

Abstract

Improvement of living standards and life expectancy due to a lot of human recombinant proteins has been developed. Many researches have tried to apply large mammals as protein producing bioreactors. However, they have got some disadvantages such as long generation duration and the difficulties in purification of recombinant proteins from a complex milk protein mixture. But chicken eggs are highlighted in this situation for their advantages. Short generation duration, high fecundity, and relative low costs for breeding support chicken eggs are excellent protein bioreactor. The most important advantage is purification of recombinant proteins in egg white is easier than in the milk. So far, most of the producing methods of transgenic chickens use viral vectors. However, the origin of the virus is almost cause of pathogenicity and their stability also results in a variety of serious issues. In this study, sperm mediated gene transfer (SMGT) to produce transgenic animals first time was established in chicken. Using Amaxa Nucleofector, the genes were transfected into the sperms that were extracted from Jeju native female chicken. After gene transfection, whole of the sperms were injected into the hens by Artificial Insemination. The eggs were harvested and incubated for egg hatching. DNA genomes of hatching chicken were extracted and performed the specific PCR for confirmation of gene transgenesis. As a results, the hatchability rate and transformation rate of chicken are 30.3 % and 63.3%, respectively. In our research, the hatchability rate and transformation rate are significantly higher in SMGT method than in viral vector method. Following this success, we will produce the transgenic chicken with specific genes such as hBMP2, cytokines that are commercial value and high effective proteins in clinical treatment. Through this study, transgenic poultry is a power bioreactor for producing a lot of human recombination proteins.