杆状病毒介导的鸡生长激素表达。

Baculovirus-mediated expression of chicken growth hormone.

作者信息

Foster D N, Proudman J A, Harmon S A, Foster L K

机构信息

Department of Animal Science, University of Minnesota, St. Paul 55108, USA.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 1997 Jun;117(2):233-9. doi: 10.1016/s0305-0491(97)00046-1.

DOI:10.1016/s0305-0491(97)00046-1

PMID:9226883

Abstract

A full-length chicken growth hormone (cGH) cDNA was placed downstream from the Autograph californica nuclear polyhedron virus, AcNPV, polyhedron gene promoter and expressed in Sf9 insect cells. Secreted recombinant cGH levels averaged 2-10 micrograms/ml from day 5-10 postinfection. The recombinant cGH analyzed by SDS-PAGE gels and Western blotting consisted of a doublet with M(r) of 26.5 and 23.5 kDa. Analysis by 2-D electrophoresis of partially-purified recombinant cGH and purified native cGH revealed similar immunoreactive charge isoforms and M(r) variants. The recombinant hormone was biologically active in a homologous radioreceptor assay. The results show that cGH expressed in insect cells is biologically and immunologically active, and that a variety of isoforms are secreted which exhibit size and charge properties similar to those of pituitary-derived cGH.

摘要

将全长鸡生长激素（cGH）cDNA置于苜蓿银纹夜蛾核型多角体病毒（AcNPV）多角体基因启动子的下游，并在Sf9昆虫细胞中表达。感染后第5至10天，分泌的重组cGH水平平均为2 - 10微克/毫升。通过SDS - PAGE凝胶和蛋白质免疫印迹分析的重组cGH由分子量为26.5 kDa和23.5 kDa的双峰组成。对部分纯化的重组cGH和纯化的天然cGH进行二维电泳分析，结果显示出相似的免疫反应性电荷异构体和分子量变体。该重组激素在同源放射受体测定中具有生物活性。结果表明，在昆虫细胞中表达的cGH具有生物学和免疫学活性，并且分泌出多种异构体，其大小和电荷特性与垂体来源的cGH相似。