Title |
The screening and functional study of proteins binding with the BmNPV polyhedrin promoter
|
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Published in |
Virology Journal, May 2012
|
DOI | 10.1186/1743-422x-9-90 |
Pubmed ID | |
Authors |
Wei Yu, Jia Li, Meihui Wang, Yanping Quan, Jian Chen, Zuoming Nie, Zhengbing Lv, Yaozhou Zhang |
Abstract |
The polyhedrin gene promoter has an essential role in regulating foreign gene expression in baculovirus expression vector systems (BEVS); however, the high-level transcription mechanism is still unknown. One-hybrid screening in yeast is a powerful way of identifying rapidly heterologous transcription factors that can interact with the polyhedrin promoter DNA sequence. In the current study, total RNA was extracted from the fat bodies of fifth-instar silkworm larvae that had been infected with Bombyx mori nuclear polyhedrosis virus (BmNPV) for 5 days; complementary DNA (cDNA) was then generated using reverse-transcription (RT)-PCR to construct a silkworm gene expression library. Key polyhedrin promoter bait sequences were synthesized to generate a bait yeast strain, which was used to screen the one-hybrid cDNA library. |
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