Kitts P A, Ayres M D, Possee R D
NERC Institute of Virology and Environmental Microbiology, Oxford, UK.
Nucleic Acids Res. 1990 Oct 11;18(19):5667-72. doi: 10.1093/nar/18.19.5667.
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.
拥有独特限制酶切位点的苜蓿银纹夜蛾多核衣壳核型多角体病毒（AcMNPV）工程衍生物提供了一种病毒DNA来源，该病毒DNA可通过用特定的核酸内切酶消化而线性化。比较了来自两种此类病毒的环状或线性化DNA的感染性和重组活性。当使用磷酸钙方法转染到草地贪夜蛾细胞中时，线性形式的感染性比相应的环状形式低15至150倍。然而，线性病毒DNA在与合适的转移载体共转染时重组能力很强。高达30%的子代病毒是重组体，这一比通过与环状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，可以促进从目前使用的任何AcMNPV转移载体中分离重组杆状病毒表达载体。
