LINEARIZATION OF BACULOVIRUS DNA ENHANCES THE RECOVERY OF RECOMBINANT VIRUS EXPRESSION VECTORS
LINEARIZATION OF BACULOVIRUS DNA ENHANCES THE RECOVERY OF RECOMBINANT VIRUS EXPRESSION VECTORS
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DOI：
10.1093/nar/18.19.5667
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发表时间：
1990-10-11
影响因子：
14.9
通讯作者：
POSSEE, RD
中科院分区：
文献类型：
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作者：
KITTS, PA;AYRES, MD;POSSEE, RD
具有独特限制性内切酶位点的海棠多核衣壳核型多角体病毒(AcMNPV)的工程化衍生物提供了一种病毒DNA的来源，可以通过特定的内切酶将其线性化。比较了两种病毒的环状DNA和线性化DNA的感染性和重组活性。用磷酸钙法将线形病毒导入果蝇细胞，其感染力比相应的圆形病毒低15-150倍。然而，线性病毒DNA擅长与适当的转移载体共转染重组。高达30%的子代病毒是重组的，比与环状AcMNPV DNA共转染获得的重组体比例高10倍。因此，通过在适当位置线性化病毒DNA，可以从目前使用的任何AcMNPV转移载体中分离重组杆状病毒表达载体。
Engineered derivatives of Autographa californica multiple nucleocapsid nuclear polyhedrosis virus (AcMNPV) possessing a unique restriction site provide a source of viral DNA that can be linearized by digestion with a specific endonuclease. Circular or linearized DNA from two such viruses were compared in terms of their infectivity and recombinogenic activities. The linear forms were 15- to 150-fold less infectious than the corresponding circular forms, when transfected into Spodoptera frugiperda cells using the calcium phosphate method. Linear viral DNA was, however, proficient at recombination on co-transfection with an appropriate transfer vector. Up to 30% of the progeny viruses were recombinant, a 10-fold higher fraction of recombinants than was obtained from co-transfections with circular AcMNPV DNA. The isolation of a recombinant baculovirus expression vector from any of the AcMNPV transfer vectors currently in use can thus be facilitated by linearization of the viral DNA at the appropriate location.