Kadono-Okuda K, Yamamoto M, Higashino Y, Taniai K, Kato Y, Chowdhury S, Xu J, Choi S K, Sugiyama M, Nakashima K
Laboratory of Biological Defense, National Institute of Sericultural and Entomological Science, Ibaraki, Japan.
Biochem Biophys Res Commun. 1995 Aug 15;213(2):389-96. doi: 10.1006/bbrc.1995.2144.
To express the cDNA encoding human growth hormone (hGH) in larvae of Bombyx mori, B. mori nuclear polyhedrosis virus (BmNPV) was employed as an expression vector. For the construction of the recombinant virus, the hGH cDNA was inserted into the downstream of the strong polyhedrin promoter to achieve a high level expression. Immunoblot analysis revealed that the virus-mediated hGH was synthesized in the larvae and secreted into the hemolymph. The yield of the recombinant hGH synthesized in the larvae reached to a level of 160 micrograms/ml of hemolymph after purification. The purified recombinant hGH was confirmed to have both the same molecular weight and amino acid sequence at its N-terminal region as those of the natural counterpart. In addition, the biological activity of the recombinant hGH was comparable to that of the natural hGH in the growth-stimulating effect on rat Nb 2 Node lymphoma cells.
为了在家蚕幼虫中表达编码人生长激素（hGH）的cDNA，使用家蚕核型多角体病毒（BmNPV）作为表达载体。为构建重组病毒，将hGH cDNA插入强多角体蛋白启动子的下游以实现高水平表达。免疫印迹分析表明，病毒介导的hGH在幼虫中合成并分泌到血淋巴中。纯化后，幼虫中合成的重组hGH产量达到血淋巴160微克/毫升的水平。经确认，纯化的重组hGH与天然hGH具有相同的分子量且其N端区域的氨基酸序列也相同。此外，重组hGH对大鼠Nb 2 淋巴瘤细胞的生长刺激作用的生物学活性与天然hGH相当。
