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斜纹夜蛾细胞系(SL 96)的建立与特性鉴定,该细胞系在19摄氏度下适应不同杆状病毒。

Establishment and characterization of Spodoptera littoralis cell line (SL 96), adapted at 19 degrees C permissive for different baculoviruses.

作者信息

Khamiss Omaima

机构信息

Center of Virology, Faculty of Agriculture-IRD, Cairo University.

出版信息

J Egypt Soc Parasitol. 2005 Dec;35(3):751-60.

Abstract

Cells from ovarioule tissues of the Egyptian cotton leaf worm, Spodoptera littoralis (SL 96) were established, identified and characterized comparing with other lepidopteran cell lines using isozymes, karyotyping, RAPD PCR, and polyacrylamide gel electrophoresis polypeptide PAGE profile. Comparison of the number of chromosomes and PAGE profiles; also the isozymes profiles demonstrate the possibility to distinguish the cell lines by those analyses and having a distinguishable finger print for every cell line. A complete replication of the S. littoralis granulovirus (SpliGV), nucleopolyherovirus (SpliNPV), and Phthorimaea operculella granulovirus (PhopGV) was obtained in vitro by both viral infection and DNA transfection in the S. littoralis (S196) cell line. Spli-GV and PhopGV multiplied extensively during several passages in S196 cells at 19 degrees C. Virus infected cells were incubated at 19 degrees C for the first 4 hours and then at .27 degrees C for the rest of the experiment (20 days). Generally, all the three viral multiplications proceeded at the same rate approximately. Comparison of Spli GV progenies multiplied either in vivo or in vitro, using electron microscopy and restriction profile analysis, showed their identity. Viral infections were detected also by specific prepared nucleic probes.

摘要

建立、鉴定并表征了埃及棉叶虫(Spodoptera littoralis,SL 96)卵巢组织的细胞,并使用同工酶、核型分析、随机扩增多态性DNA聚合酶链式反应(RAPD PCR)以及聚丙烯酰胺凝胶电泳多肽图谱(PAGE图谱),与其他鳞翅目细胞系进行比较。对染色体数量和PAGE图谱进行比较;同工酶图谱也表明,通过这些分析有可能区分细胞系,并且每个细胞系都有可区分的指纹图谱。通过病毒感染和DNA转染,在埃及棉叶虫(S196)细胞系中实现了埃及棉叶虫颗粒体病毒(SpliGV)、核型多角体病毒(SpliNPV)和马铃薯块茎蛾颗粒体病毒(PhopGV)的体外完全复制。Spli - GV和PhopGV在19℃下于S196细胞中传代培养多次后大量增殖。病毒感染的细胞在19℃下孵育4小时,然后在实验剩余时间(20天)内于27℃下孵育。一般来说,所有三种病毒的增殖速度大致相同。使用电子显微镜和限制性图谱分析比较了在体内或体外增殖的Spli GV子代,结果显示它们具有一致性。还通过特异性制备的核酸探针检测到病毒感染。

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