家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)是一种具有较强传染性的病毒,在感染的极晚期超高水平表达一种约29 kD的碱溶性多角体蛋白。但该蛋白为病毒复制的非必需蛋白,因此,利用该基因可以被删除或被外源基因替换而...家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)是一种具有较强传染性的病毒,在感染的极晚期超高水平表达一种约29 kD的碱溶性多角体蛋白。但该蛋白为病毒复制的非必需蛋白,因此,利用该基因可以被删除或被外源基因替换而不影响病毒的复制的原理构建重组病毒,可以实现外源基因的高水平表达。然而,目前虽已经有较多文献研究多角体蛋白高水平表达的分子机制,但迄今对于多角体蛋白基因编码区的结合蛋白尚不明确。本研究通过DNA pull-down联合质谱技术对结合在多角体蛋白编码区的蛋白质进行鉴定,共鉴定到78个宿主来源的蛋白和49个病毒自身编码的蛋白。对这些蛋白进行GO注释,结果表明这些蛋白除了具有共同的核酸结合功能外,还有其他不同的生物学功能,共同在多角体蛋白的超高水平表达过程中发挥作用。进一步从上述蛋白中筛选出了最值得深入研究的9个宿主蛋白和10个病毒蛋白。对这些蛋白进行深入研究,将为深入揭示多角体蛋白超高水平表达的分子机制提供新的线索。展开更多
Hyphantria cuea nucleopolyhedrovirus(HcNPV) and several other nucleopolyhedrosis was extracted by differential centrifugation,and purified with sucrose gradient centrifugation method.Polyhedrin was gotten and analysed...Hyphantria cuea nucleopolyhedrovirus(HcNPV) and several other nucleopolyhedrosis was extracted by differential centrifugation,and purified with sucrose gradient centrifugation method.Polyhedrin was gotten and analysed by SDS-PAGE.It indicated that most of those proteins had 32 KD protein band,and with 64 KD protein,and might be the main protein dimer with the structure and found Hyphantria cunea nuclear polyhedrosis virus protein in 18 KD department had a specific band.展开更多
The polyhedrin gene of EpNPV was located on Hin d III E, Xba I B, Eco RI J, Kpn I A, Pst I F, Bam H I G fragments under stringent condition(68℃,in water),by using the DIG labeled Bam H I F fragment of AcMNPV DNA as t...The polyhedrin gene of EpNPV was located on Hin d III E, Xba I B, Eco RI J, Kpn I A, Pst I F, Bam H I G fragments under stringent condition(68℃,in water),by using the DIG labeled Bam H I F fragment of AcMNPV DNA as the probe.In order to sequence the EpNPV polyhedrin gene,the Bam H I G fragment was cloned into pTZ19R vector.Then the fragment was digested by Xba I? Hin d III,and subcloned into pTZ19R.展开更多
文摘家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)是一种具有较强传染性的病毒,在感染的极晚期超高水平表达一种约29 kD的碱溶性多角体蛋白。但该蛋白为病毒复制的非必需蛋白,因此,利用该基因可以被删除或被外源基因替换而不影响病毒的复制的原理构建重组病毒,可以实现外源基因的高水平表达。然而,目前虽已经有较多文献研究多角体蛋白高水平表达的分子机制,但迄今对于多角体蛋白基因编码区的结合蛋白尚不明确。本研究通过DNA pull-down联合质谱技术对结合在多角体蛋白编码区的蛋白质进行鉴定,共鉴定到78个宿主来源的蛋白和49个病毒自身编码的蛋白。对这些蛋白进行GO注释,结果表明这些蛋白除了具有共同的核酸结合功能外,还有其他不同的生物学功能,共同在多角体蛋白的超高水平表达过程中发挥作用。进一步从上述蛋白中筛选出了最值得深入研究的9个宿主蛋白和10个病毒蛋白。对这些蛋白进行深入研究,将为深入揭示多角体蛋白超高水平表达的分子机制提供新的线索。
文摘Hyphantria cuea nucleopolyhedrovirus(HcNPV) and several other nucleopolyhedrosis was extracted by differential centrifugation,and purified with sucrose gradient centrifugation method.Polyhedrin was gotten and analysed by SDS-PAGE.It indicated that most of those proteins had 32 KD protein band,and with 64 KD protein,and might be the main protein dimer with the structure and found Hyphantria cunea nuclear polyhedrosis virus protein in 18 KD department had a specific band.
文摘The polyhedrin gene of EpNPV was located on Hin d III E, Xba I B, Eco RI J, Kpn I A, Pst I F, Bam H I G fragments under stringent condition(68℃,in water),by using the DIG labeled Bam H I F fragment of AcMNPV DNA as the probe.In order to sequence the EpNPV polyhedrin gene,the Bam H I G fragment was cloned into pTZ19R vector.Then the fragment was digested by Xba I? Hin d III,and subcloned into pTZ19R.
