@article{MAKHILLJAVA201211163649,
    title = {Expression of Human Granulocyte-Macrophage Colony-Stimulating Factor in Stably-Transformed BmN and Sf-9 Cells and Silkworms by a Non-Transposon Vector},
    journal = {Journal of Animal and Veterinary Advances},
    volume = {11},
    number = {16},
    pages = {2890-2897},
    year = {2012},
    issn = {1680-5593},
    doi = {javaa.2012.2890.2897},
    url = {https://makhillpublications.co/view-article.php?issn=1680-5593&doi=javaa.2012.2890.2897},
    author = {Cheng Liang,Hao Kun,Guang Li,Yan Mei and},
    keywords = {Non-transposon vector,BmN cells,Sf-9 cells,Bombyx mori,hGM-CSF,transgene},
    abstract = {This study aimed to explore the possibility of non-transposon vector mediated foreign gene expression in cultured insect cells and transgenic silk worms. To this end, the human Granulocyte-Macrophage Colony-Stimulating Factor (<I>hGM-CSF</I>) gene was inserted into the insect cell expression vector pIZT-V5-His to generate the recombinant vector pIZT-hGM-CSF. After transfection of BmN and Sf-9 cells with the pIZT-hGM-CSF vector, stably-transformed cells expressing the <I>hGM-CSF</I> gene were selected using the antibiotic zeocin at a final concentration of 300-400 &#956;g mL<SUP>-1</SUP>. Expression of a 22 kDa protein band representing hGM-CSF was detected in the transformed cells by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. The expression levels of hGM-CSF in BmN and Sf-9 cells were determined by Enzyme-Linked Immunosorbent Assay (ELISA) to be about 0.7 and 0.3 ng/10<SUP>6</SUP> cells, respectively. The transgenic vector pIZT-hGM-CSF was transferred into silkworm eggs using sperm-mediated gene transfer. Transgenic silkworms were obtained after screening for the <I>gfp </I>gene and were verified by polymerase chain reaction, dot hybridization and Western blotting. The expression level of hGM-CSF determined by ELISA was about 4.7 ng g<SUP>-1</SUP> of freeze-dried silk glands in the G5 generation. These results suggest that heterologous genes can be integrated into cultured BmN and Sf-9 cells and into the silkworm genome using a non-transposon vector and can be expressed successfully.}
    }