Expression of Marek,s Disease Virus (MDV) Gene Homologous to UL9 Gene of Herpes Simplex Virus Type 1(HSV-1) by Recombinant Baculovirus

CNU IR > Chna Nan Annual Bulletin > Vol.25 (1999) > Item 310902800/22534

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Title:	Expression of Marek,s Disease Virus (MDV) Gene Homologous to UL9 Gene of Herpes Simplex Virus Type 1(HSV-1) by Recombinant Baculovirus 以重組昆蟲桿狀病毒表現馬雷克病毒之UL9蛋白質
Authors:	Ting-Feng wu Jeng-Chai Hwang Hong-Hwa Chen
Contributors:	藥學系
Keywords:	Marek's disease virus Marek's disease virus UL9protein homologous recombination 馬雷克病毒馬雷克UL9蛋白質同源重組
Date:	1999
Issue Date:	2010-04-08 13:04:55 (UTC+8)
Abstract:	In previous studies, a MDV UL9 homologue gene was identified which lies between the BamHI C and BamHI G fragments of the serotype 1 MDV strain GA genome. Computer analysis of the predicted amino acid sequences of MDV and HSV-1 UL9 proteins revealed that MDV UL9 is highly similar to HSV-1 UL9. MDV and HSV-1 UL9 proteins share numerous structural motifs, suggesting that MDV UL9 may have biochemical activities similar to HSV-1 UL9, specifically the origin-binding activity. MDV UL9 protein was shown to bind to the origin of MDV DNA replication by using electrophoretic mobility gel shifts (EMSAs). EMSAs was performed with in vitro expressed MDV UL9 protein or with nuclear extracts prepared from Chicken embryonic fibroblast (CEF)cells infected with serotype 1 MDV. The probes used in located within the origin of MDV DNA replication. In this study, the recombinant beculovirus will be constructed by homologous recombination. The recombinant baculovirvs will be constructed by homologous recombination. The recombinant baculovirus contains the MDV UL9 gene under the control OT polyhedrin promoter. In the future, MDV UL9 protein will be purified from Sf9 cells infected by the recombinant baculovirus and used in detecting other biochemical activities of MDV UL9 protein. 在先前的研究裡，我們已確認馬達克病毒之UL9基因位於血清一型(serotype 1)之馬克病毒基因體(genome)的BamHI C與BamHI G片段之間，依照預測的胺基酸順序之電腦分析結果顯示，馬雷克病毒的UL9基因與第一型q疱疹病毒(herpes simplex virus type 1 )之UL9基因有許多相似之構造單位(structural motifs)。所以馬雷克病毒之UL9也許擁有與庖疹病毒之UL9基因相似之生1t功能，特別是複製起始點(origin)的附著能力。另外，我們使用活體外產生之馬雷克病毒UL9蛋白質或由受馬雷克病毒感染之細胞抽出的核萃取物進行膠體位移實驗，證實馬雷克病毒UL9蛋白可與馬雷克病毒之複始點結合。在此研究裡，我們將利用同源重組(homologous rcombination)原理製備含有馬雷克病毒UL9基因之昆蟲桿狀病毒(Baculovirus),在重組昆蟲桿狀病毒中，馬雷克病毒UL9基因由核多角體蛋白質基因(polyhedrin gene)之起動子驅動。含有馬克病毒UL9基因之昆蟲桿狀病毒將來可被利用來感染Sf9細胞，並藉以純化馬雷克病毒UL9蛋白質，以便將來用於測定馬雷克病毒UL9蛋白質的其他生化活性。
Relation:	嘉南學報 25 : p.165-177
Appears in Collections:	[Chna Nan Annual Bulletin] Vol.25 (1999) [Dept. of Pharmacy] Periodical Articles

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