关键词: 人组织型纤溶酶原激活剂, 卵黄高磷蛋白, 功能性食品, 卵黄, 转基因

Abstract:

In order to explore the translocation feasibility of human tissue-type plasminogen activator (t-PA) from liver to egg yolk and develop functional eggs with anti-cardiovascular diseases, a recombinant plasmid pPhosvitin-tPA encoded human tissue-type plasminogen activator was constructed. The pcDNA3-tPA and pPhosvitin-tPA coated with liposome was directly injected into hen s liver. The expression of recombinant t-PA protein in liver and egg yolk was evaluated using Western blot and ELISA, and its biological activity in egg yolk was determined using fibrinolysis test. SDS-PAGE and Western blot confirmed that recombinant t-PA protein was expressed in yolk on the 7th day post-injection; the molecular weight of the expressed protein was approximately 63 kD. The expression rates were 39.16 mg/L and 53.92 mg/L measured in ELISA. Fibrinolysis of t-PA in vitro indicated that the expressed protein had the capability to activate tissue-type plasminogen. Therefore, it was a feasible strategy to obtain transgentic eggs with recombinant t-PA through its expression in hen s liver subjected to the injection of plasmid encoded recombinant t-PA, which provided a novel method to develop functional food with anti-cardiovascular diseases and made it reality to become a new resource of protein drugs from hens.

Key words: human tissue-type plasminogen activator (t-PA), phosvitin, functional food, yolk, transgenic egg