To generate large amounts of a recombinant protein, many investigators have turned to baculovirus expression systems. Compared to bacterial expression systems, the posttranslational processing and folding of recombinant proteins produced in insect cells more closely resembles mammalian processes, and the yields of functional protein are often much greater.
To generate large amounts of a recombinant protein, many investigators have turned to baculovirus expression systems. Compared to bacterial expression systems, the posttranslational processing and folding of recombinant proteins produced in insect cells more closely resembles mammalian processes, and the yields of functional protein are often much greater.
BacPAK method
The BacPAK System uses the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) to produce target proteins in insect cells. The target gene is inserted into a shuttle vector, which is cotransfected into insect host cells with the linearized BacPAK6 Viral DNA. The specially designed BacPAK6 Viral DNA forces recombination between the virus and transfer vector, resulting in high recombination efficiency. Following recombination, a few viral plaques are picked and purified, and the recombinant phenotype is verified. The newly isolated recombinant virus can then be amplified and used to infect insect cell cultures to produce large amounts of the desired protein.If you wish, you can use pBacPAK8-GUS (sold as part of the BacPAK Baculovirus Expression System) as a positive control for the cotransfection step. This transfer vector has the E. coli beta-glucuronidase (GUS) gene cloned downstream of its polyhedrin promoter. Recombination of pBacPAK8-GUS with the BacPAK6 DNA digest generates recombinant viruses that express beta-glucuronidase. Expression of GUS can be detected by generation of a blue color from the chromogenic GUS substrate X-GLUC.
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这三样药是能混合在一起吃？
谢谢。
深夜，保姆觉得无聊就想去楼下看电视。但是她看不了，因为楼下没有电视（因为孩子的父母不希望他们的孩子看太多垃圾）。她就打电话给孩子的父母，问是否可以在他们的卧室看电视，当然孩子的父母同意了。
但保姆又想要最后一个请求。
她问是否可以用毯子或者衣服盖住那小丑雕像，因为那使她感到很害怕。
电话沉默了一会。
（此时爸爸在和保姆通话）
他说：带孩子离开房间……
我们将会叫警察……我们从来没有什么小丑雕像。
那小丑很可能是一个从监狱逃出来的杀人犯。
电话里沉默了一会儿。
（正在跟保姆通话的孩子的父亲）说：带上孩子们，离开房子……我们会通知警察……我们没有一个小丑雕像……
孩子们和保姆被小丑谋杀了。
结果是，小丑是一个从监狱里逃出来的杀人犯。
如果你不在5分钟内转发这个贴子，这个小丑在凌晨3点时将会拿着刀站在你的床前。
我在这里发了，这就是恶魔般的小丑没有杀我的原因
