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鹦鹉热衣原体主要外膜蛋白在COS细胞和火鸡骨骼肌中的高水平表达。

High-level expression of Chlamydia psittaci major outer membrane protein in COS cells and in skeletal muscles of turkeys.

作者信息

Vanrompay D, Cox E, Mast J, Goddeeris B, Volckaert G

机构信息

Laboratory of Gene Technology, University of Leuven, 3001 Leuven, Belgium.

出版信息

Infect Immun. 1998 Nov;66(11):5494-500. doi: 10.1128/IAI.66.11.5494-5500.1998.

Abstract

The omp1 genes encoding the major outer membrane proteins (MOMPs) of avian Chlamydia psittaci serovar A and D strains were cloned and sequenced. The nucleotide sequences of the avian C. psittaci serovar A and D MOMP genes were found to be 98.9 and 87.8% identical, respectively, to that of the avian C. psittaci serovar A strain 6BC, 84.6 and 99.8% identical to that of the avian C. psittaci serovar D strain NJ1, 79.1 and 81.1% identical to that of the C. psittaci guinea pig inclusion conjunctivitis strain, 60.9 and 62.5% identical to that of the Chlamydia trachomatis L2 strain, and 57.5 and 60.4% identical to that of the Chlamydia pneumoniae IOL-207 strain. The serovar A or D MOMPs were cloned in the mammalian expression plasmid pcDNA1. When pcDNA1/MOMP A or pcDNA1/MOMP D was introduced into COS7 cells, a 40-kDa protein that was identical in size, antigenicity, and electrophoretic mobility to native MOMP was produced. Recombinant MOMP (rMOMP) was located in the cytoplasm of transfected COS7 cells as well as in the plasma membrane and was immunoaccessible. Intramuscular administration of pcDNA1/MOMP in specific-pathogen-free turkeys resulted in local expression of rMOMP in its native conformation, after which anti-MOMP antibodies appeared in the serum.

摘要

对编码禽鹦鹉热衣原体A和D血清型菌株主要外膜蛋白(MOMP)的omp1基因进行了克隆和测序。发现禽鹦鹉热衣原体A和D血清型MOMP基因的核苷酸序列分别与禽鹦鹉热衣原体A血清型6BC菌株的核苷酸序列有98.9%和87.8%的同一性,与禽鹦鹉热衣原体D血清型NJ1菌株的核苷酸序列有84.6%和99.8%的同一性,与鹦鹉热衣原体豚鼠包涵体结膜炎菌株的核苷酸序列有79.1%和81.1%的同一性,与沙眼衣原体L2菌株的核苷酸序列有60.9%和62.5%的同一性,与肺炎衣原体IOL-207菌株的核苷酸序列有57.5%和60.4%的同一性。血清型A或D的MOMP被克隆到哺乳动物表达质粒pcDNA1中。当将pcDNA1/MOMP A或pcDNA1/MOMP D导入COS7细胞时,产生了一种大小、抗原性和电泳迁移率与天然MOMP相同的40 kDa蛋白。重组MOMP(rMOMP)位于转染的COS7细胞的细胞质以及质膜中,并且具有免疫可及性。在无特定病原体的火鸡中肌肉注射pcDNA1/MOMP导致rMOMP以其天然构象在局部表达,之后血清中出现了抗MOMP抗体。

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