FUNCTIONAL ANALYSIS OF DENGUE NONSTRUCTURAL PROTEINS, NS3 AND NS5

登革病毒NS3和NS5非结构蛋白是候选者 对于参与病毒RNA转录的病毒酶 和/或复制。 为了检测基因表达， 为了研究这两种蛋白质的功能活性，我们改造了NS3， NS5特异性DNA，并构建了重组杆状病毒， 表达NS3或NS5病毒基因。 NS3的合成 间接免疫荧光法检测重组病毒感染的细胞 免疫荧光测定。 类似地，NS5的合成是 通过SDS-聚丙烯酰胺凝胶分析来自 重组病毒感染细胞。 蛋白质产品是 制备用于兔免疫的抗血清和对这些特异性的抗血清 获得蛋白质。 NS3特异性血清的可用性 应该有助于在设计的实验中检测这种蛋白质 研究下游NS3a和NS4b序列的需求， NS3的表达。 这些实验将进行测试 假设NS1和NS3可能是类似的，因为每个都是 接着是两个可以作为蛋白酶的疏水蛋白 其影响登革多蛋白的特异性切割。

The dengue virus NS3 and NS5 nonstructural proteins are candidates for viral enzymes that are involved in viral RNA transcription and/or replication. In order to examine the gene expression and functional activity of these two proteins, we engineered NS3 and NS5 specific DNA and constructed a recombinant baculovirus which expressed the NS3 or NS5 viral gene. Synthesis of NS3 in recombinant virus-infected cells was detected by indirect immunofluorescence assay. Similarly, the synthesis of NS5 was detected by SDS-polyacrylamide gel analysis of a lysate from recombinant virus infected cells. The protein products were prepared for immunization of rabbits and antisera specific to these proteins were obtained. The availability of NS3 specific sera should facilitate detection of this protein in experiments designed to study the requirement of downstream NS3a and NS4b sequences for expression of NS3. These experiments will be carried out to test the hypothesis that NS1 and NS3 may be analogous since each is followed by two hydrophobic proteins that may serve as proteases that effect specific cleavage of the dengue polyprotein.