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Expression of hGM-CSF in silk glands of transgenic silkworms using gene targeting vector

文献类型： 外文期刊

作者： Xue, Renyu 1 ; Chen, Huimei 2 ; Cui, Linlin 2 ; Cao, Guangli 1 ; Zhou, Wenlin 3 ; Zheng, Xiaojian 2 ; Gong, Chengliang 1 ;

作者机构： 1.Soochow Univ, Natl Engn Lab Modern Silk, Suzhou 215123, Peoples R China

2.Soochow Univ, Preclin Med & Biol Sci Coll, Suzhou 215123, Peoples R China

3.Zhejiang Acad Agr Sci, Sericulture Res Inst, Hangzhou 310021, Zhejiang, Peoples R China

关键词： Bombyx mori;Gene targeting;Silk gland;hGM-CSF

期刊名称：TRANSGENIC RESEARCH （ 影响因子：2.788； 五年影响因子：2.377 ）

ISSN： 0962-8819

年卷期： 2012 年 21 卷 1 期

页码：

收录情况： SCI

摘要： The silk gland of the silkworm is a highly specialized organ that has the wonderful ability to synthesize and secrete silk protein. To express human granucyto-macrophage colony-stimulating factor (hGM-CSF) in the posterior silk glands of gene-targeted silkworms, a targeting vector pSK-FibL-L-A3GFP- PH-GMCSF-LPA-FibL-R was constructed, harboring a 1.2 kb portion of the left homogenous arm (FibL-L), a 0.5 kb portion of the right homogenous arm (FibL-R), fibroin H-chain-promoter-driven hGM-CSF and silkworm actin 3-promoter-driven gfp. The targeting vector was then introduced into the eggs of silkworm, and the transgenic silkworms were verified by PCR and DNA hybridization after being screened for the gfp gene. Western blotting analysis using an antibody against hGM-CSF demonstrated a specific band with a molecular weight of 22 kD in the silk glands of the G3 generation transgenic silkworms. The level of expression of hGM-CSF in the posterior silk glands of the G3 generation transgenic silkworms was approximately 2.70 ng/g of freezedried powdered posterior silk gland. These results showed that the heterologous gene could be introduced into the silkworm genome and expressed successfully. Further more, the exogenous genes existing in the G5 transgenic silkworm identified by PCR confirmed its integration stability. In addition, the silk glands containing expressed hGM-CSF performed the function of significantly increasing leukocyte count of CY-treated mice in a time-and-dose-dependent manner.

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