β-半乳糖苷酶和荧光素酶在感染重组苜蓿银纹夜蛾核型多角体病毒的昆虫细胞系中的表达

Expression of beta-galactosidase and luciferase in insect cell lines infected with a recombinant AcMNPV.

作者信息

McIntosh A H, Grasela J J

机构信息

USDA, ARS Biological Control of Insects Research Laboratory, Columbia, Missouri 65205.

出版信息

In Vitro Cell Dev Biol Anim. 1994 Apr;30A(4):275-8. doi: 10.1007/BF02632051.

DOI:10.1007/BF02632051

PMID:8069450

Abstract

An AcMNPV recombinant (Ac-gal-luc) carrying the beta-galactosidase and luciferase genes under the control of the p10 and polyhedrin promoters, respectively, was used to study expression in nine insect cell lines. All AcMNPV-permissive cell lines expressed both reporter genes with the coleopteran cell line, Anthonomus grandis (AGE), producing the highest concentrations of beta-galactosidase (5.0 x 10(6) pg/ml) and luciferase (2.67 x 10(3) pg/ml). Both enzymes were detected as early as 12 h postinoculation in lysate samples of the AGE cell line. Helicoverpa armigera (HA), a nonpermissive cell line, expressed beta-galactosidase at 72 h postinoculation at a concentration of 3.5 x 10(3) pg/ml. However, expression of luciferase was not detected. Expression of luciferase and beta-galactosidase was also not detected in the nonpermissive Helicoverpa zea (HZ) cell line.

摘要

一种携带β-半乳糖苷酶和荧光素酶基因的苜蓿银纹夜蛾核型多角体病毒重组体（Ac-gal-luc），分别在p10和多角体蛋白启动子的控制下，被用于研究在9种昆虫细胞系中的表达情况。所有对苜蓿银纹夜蛾核型多角体病毒敏感的细胞系都表达了这两种报告基因，其中鞘翅目昆虫细胞系大谷实蝇（AGE）产生的β-半乳糖苷酶（5.0×10⁶ pg/ml）和荧光素酶（2.67×10³ pg/ml）浓度最高。在接种后12小时，在AGE细胞系的裂解物样本中就检测到了这两种酶。非敏感细胞系棉铃虫（HA）在接种后72小时表达了β-半乳糖苷酶，浓度为3.5×10³ pg/ml。然而，未检测到荧光素酶的表达。在非敏感细胞系玉米螟（HZ）中也未检测到荧光素酶和β-半乳糖苷酶的表达。