利用桿狀病毒載體系統研究在昆蟲細胞中B型肝炎表面抗原之合成與分泌作用

Abstract

以市售桿狀病毒轉形試劑組所得之重組病毒比例，並非如預期的接近百分之百，因此擬得純系化之重組病毒及較高之重組蛋白產量，至少須經兩次以上病毒純化。接著，取純化過並載有B型肝炎表面抗原M protein基因之重組病毒，與兩株僅載有S protein基因之重組病毒分別感染昆蟲細胞，發現S protein幾乎不分泌至培養液，絕大部分累積在細胞內；反觀在感染後四十八小時即於培養液中偵測到相當量之分泌性M protein。此外，當昆蟲細胞中同時存在M與S protein時，M protein將有抑制S protein分泌之可能。

The fraction of recombinant virus which was obtained from cotransfection fluids using commercial transfection kit was not nearly 100% as expected.So purified recombinants and higher recombinant protein production could only be approached when undergone virus purification twice.Then we analyzed the secretion of HBV M and S proteins from three recombinant viruses containing either M or S protein gene. The results indicated that the S proteins were predominantly accumulated in the insect cells and were inefficiently secreted into the medium.In contrast, there were comparatively high level of the M proteins in the medium two days after infection.On the other hand, the secretion of the S proteins were presumably inhibited by the presence of the M proteins.