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10型禽腺病毒抗原在大肠杆菌中的表达

Expression of fowl adenovirus type 10 antigens in Escherichia coli.

作者信息

Sheppard M, Werner W

机构信息

CSIRO, Animal Health Research Laboratory, Parkville, Victoria, Australia.

出版信息

Vet Microbiol. 1990 Aug;24(2):105-12. doi: 10.1016/0378-1135(90)90057-3.

Abstract

In an attempt to construct a genetic map of the fowl adenovirus (FAV) and to determine which viral proteins are natural immunogens in chickens and hence may be relevant to protective immunity we have constructed an expression library of FAV type 10 DNA. The genomic DNA was partially digested with the restriction endonuclease Sau3A, and this DNA was inserted into the 3' terminal end of the beta-galactosidase gene in a plasmid vector. To date, approximately 600 clones have been identified that express FAV type 10 antigens as determined by immunological screening with rabbit antisera to purified virus, including one that has amino acid homology with the 100 kDa protein of human adenovirus type 5. These antigen positive clones were found to contain DNA from FAV type 10 genome as determined by hybridisation to FAV DNA. These clones will allow the further characterisation of FAV and possibly the identification of potential vaccine molecules.

摘要

为构建禽腺病毒(FAV)的遗传图谱,并确定哪些病毒蛋白是鸡体内的天然免疫原,因此可能与保护性免疫相关,我们构建了10型FAV DNA的表达文库。基因组DNA用限制性内切酶Sau3A进行部分消化,然后将该DNA插入质粒载体中β-半乳糖苷酶基因的3'末端。迄今为止,通过用抗纯化病毒的兔抗血清进行免疫筛选,已鉴定出约600个表达10型FAV抗原的克隆,其中一个与5型人腺病毒的100 kDa蛋白具有氨基酸同源性。通过与FAV DNA杂交确定,这些抗原阳性克隆含有来自10型FAV基因组的DNA。这些克隆将有助于对FAV进行进一步的特性分析,并有可能鉴定出潜在的疫苗分子。

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