Increase of host cell longevity by the expression of 30K protein originating from silkworm hemolymph in an insect cell-baculovirus system
Increase of host cell longevity by the expression of 30K protein originating from silkworm hemolymph in an insect cell-baculovirus system
复制标题
DOI：
10.1016/j.enzmictec.2004.08.024
复制
发表时间：
2004-12-01
影响因子：
3.4
通讯作者：
Park, TH
中科院分区：
文献类型：
--
作者：
Kim, EJ;Park, TH
将家蚕血淋巴30 K蛋白基因与野生型AcMNPV DNA进行同源重组，构建了重组杆状病毒。用重组杆状病毒（AcMNPV/30 K）感染Sf 9细胞，成功表达了30 K蛋白。为了研究30 K蛋白表达产生的效果，将感染后的宿主细胞活力与用AcMNPV/β-gal感染的Sf 9细胞的活力进行比较。感染AcMNPV/beta-gal后3d细胞活力开始指数下降，而感染AcMNPV/30 K后5d细胞活力仍维持在较高水平。这表明30 K蛋白增加了病毒感染后的细胞寿命。这种增加的细胞寿命被认为是由于杆状病毒诱导的宿主细胞凋亡的抑制，并且通过流式细胞术方法测量凋亡的程度。30 K蛋白的表达降低了亚G1级分的百分比，亚G1级分代表细胞凋亡的程度。这表明昆虫细胞中30 K蛋白的表达通过抑制细胞凋亡增加宿主细胞寿命。(C)2004爱思唯尔公司All rights reserved.
A recombinant baculovirus was constructed by the homologous recombination between wild-type AcMNPV DNA and a baculovirus transfer vector containing a gene coding for the 30K protein originating from silkworm hemolymph. The 30K protein was successfully expressed in Sf9 cells infected with the recombinant baculovirus (AcMNPV/30K). To investigate the effect produced by the expression of the 30K protein, host cell viability after infection was compared with that of Sf9 cells infected with AcMNPV/beta-gal. The viability of the cells infected with AcMNPV/beta-gal began to decrease exponentially 3 days after infection, whereas that of the cells infected with AcMNPV/30K remained at a high level until 5 days after infection. This indicates that the 30K protein increases cell longevity after viral infection. This increased cell longevity is considered to be due to the inhibition of host cell apoptosis induced by a baculovirus, and the extent of apoptosis was measured by the flow cytometric method. The percentage of the sub-G1 fraction, which represents the extent of apoptosis, was decreased by the expression of the 30K protein. This indicates that the expression of the 30K protein in insect cells increases host cell longevity by inhibiting apoptosis. (C) 2004 Elsevier Inc. All rights reserved.