Effect of silkworm hemolymph on Escherichia coli β-galactosidase expression in insect cell lines infected with a recombinant baculovirus
Effect of silkworm hemolymph on Escherichia coli β-galactosidase expression in insect cell lines infected with a recombinant baculovirus
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蚕血淋巴对重组杆状病毒感染昆虫细胞系中大肠杆菌β-半乳糖苷酶表达的影响
DOI：
10.11416/kontyushigen1930.64.477
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
T. Kawarabata
中科院分区：
文献类型：
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作者：
T. Hara;H. Tarui;H. Kawaguchi;N. Yoshimura;T. Kawarabata
以重组杆状病毒AC360为载体，用大肠杆菌/3-半乳糖苷酶基因替换卡氏拟革兰氏菌核型多角体病毒的多角体蛋白基因，检测了重组杆状病毒感染昆虫细胞后的/3-半乳糖苷酶(13-Gal)的表达。检测了多达5个据信来自家蚕的细胞系，仅在命名为SPC BM 36的细胞系中表达了对应于3-Gal的120 kDa蛋白。在感染的SPC BM 3.6细胞中，13-Gal的表达量随着添加TC-100的家白杆菌血淋巴浓度的增加而增加。在血淋巴的2%时，表达的/3-Gal蛋白占细胞总蛋白的5.4%，相当于每5.0×105个细胞的细胞内13-Gal活性的7.5×103个单位，相当于在含10%胎牛血清的TC-100培养液中的表达。因此，家蚕血淋巴对重组杆状病毒在TC-100培养的SPC BM 36细胞中表达外源基因有一定的促进作用。九州大学农学部，福冈市东湖箱崎市812号。
Expression of/3-galactosidase(13-Gal) was examined in insect cell lines infected with a recombinant baculovirus, Ac360, in which polyhedrin gene of Autographa californica nuclear polyhedrosis virus was replaced by Escherichia coli/3-Gal gene. Up to 5 cell lines believed to be derived from Bombyx mori were examined and a 120 kDa-protein corresponding to, 3-Gal was expressed only in the cell line named SPC Bm 36. The amount of 13-Gal expression in infected SPC Bm 3.6 cells increased with increasing concentration of B. mori hemolymph supplemented to the TC-100 medium. At 2% of the hemolymph, the amount of expressed/3-Gal protein accounted for 5.4% of the total cellular proteins, or 7.5 x 103 units of intracellular 13-Gal activity per 5.0 x 105 cells, which was equivalent to that expressed in the TC-100 medium with 10% fetal bovine serum. It is thus suggested that B. mori hemolymph has beneficial effects on the expression of foreign genes by recombinant baculoviruses in SPC Bm 36 cells cultured in the TC-100 medium. Faculty of Agriculture, Kyushu University, Hakozaki, Higashi-hu, Fukuoka 812.