Sequence analysis and prokaryotic expression of HearNPV ORF44
2007
Li Meng | Zhang Yalin | Wang Dun
Chinese. 棉铃虫核型多角体病毒 (HearNPV) ORF44是一个功能未知的基因，研究对HearNPV ORF44基因的序列及原核表达进行了初步研究。结果表明，HearNPV ORF44基因全长1 137 nt，编码378aa，预测编码蛋白分子量为42.7 ku。序列分析显示，ORF44具有双相启动子特征序列，同时兼有杆状病毒早期和晚期基因转录启动子特征序列。ORF44的推导氨基酸序列中含有1个强跨膜螺旋区域，32个磷酸化位点和4个糖基化位点。除HzNPV ORF45外，未发现与ORF44氨基酸同源性高于30%的基因。构建了pET-44表达载体，并在大肠杆菌BL21中得到了高效表达，表达的融合蛋白分子量为44 ku，与预测编码蛋白分子量大小相符。序列分析结果表明，ORF44是HearNPV的一个特有基因，ORF44融合蛋白的成功表达为深入研究HearNPV ORF44奠定了基础。
Show more [+] Less [-]English. The function of HearNPV ORF44 is unknown for the present, thus the HearNPV ORF44 was studied and the primary results were reported in this article. HearNPV ORF44 is 1 137 nt long, encoding a putative protein of 378aa with a predicted size of 42.7 ku. Both baculovirus consensus early and late promoter motifs are found in ORF44. One transmembrane helices region, 32 putative phosphorylation sites and 4 N-linked glycosylation sites are found in the predicted ORF44 protein. No homologue that shares an identity over 30% through NPVs and GVs is found in GenBank and SWISS-PROT. A pET-44 plasmid is constructed and ORF44 is expressed in the Eschechia coli (BL21). The molecular weight of expressed fusion protein is 44 ku, which conforms to the predicted size. The results show that ORF44 is unique to HearNPV among NPVs, and the expression of fused ORF44 protein provides the basic information on the analysis of HearNPV ORF44 function.
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