Secretion of recombinant rat annexin 5 by insect cells in a baculovirus expression system.
Secretion of recombinant rat annexin 5 by insect cells in a baculovirus expression system.
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昆虫细胞在杆状病毒表达系统中分泌重组大鼠膜联蛋白 5。
DOI：
10.1006/bbrc.1994.1609
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发表时间：
1994
影响因子：
3.1
通讯作者：
M. Kawaminami
中科院分区：
文献类型：
--
作者：
Kazuaki Takehara;S. Uchida;N. Marumoto;T. Asawa;S. Osugi;S. Kurusu;I. Hashimoto;M. Kawaminami
大鼠膜联蛋白5在昆虫细胞中表达使用杆状病毒载体，苜蓿银纹夜蛾核型多角体病毒。大鼠膜联蛋白5 cDNA的制备通过聚合酶链反应使用mRNA从大鼠垂体和多角体蛋白启动子的控制下。基因产物为36 k道尔顿，并被抗大鼠膜联蛋白5血清识别。证实了重组膜联蛋白5与膜的钙依赖性结合。接种后21小时，重组蛋白大量出现在培养基中，这是该重组病毒所特有的。高钾环境（20 mM KCl）2小时增加了重组蛋白的释放，但对于为对照制备的重组β-半乳糖苷酶没有增加。这些结果表明，缺乏信号序列的膜联蛋白5基因的产物在昆虫细胞中遵循分泌途径。
Rat annexin 5 was expressed in insect cells using a baculovirus vector, Autographa californica nuclear polyhedrosis virus. The rat annexin 5 cDNA was prepared by a polymerase chain reaction using mRNA from rat pituitary glands and placed under the control of the polyhedrin promoter. The gene product was 36 k dalton and was recognized by anti-rat annexin 5 serum. The calcium dependent binding of the recombinant annexin 5 to membranes was confirmed. The recombinant protein appeared in the medium by 21 hours post-inoculation in high amount and this was specific to this recombinant virus. High potassium milieu (20 mM KCl) for two hours increased the release of the recombinant protein but not for the recombinant beta-galactosidase prepared for a control. These results reveal that the product of the annexin 5 gene, which lacks a signal sequence, follows a secretory pathway in insect cells.