%0 Journal Article
%T Construction of Marek''''s Disease Virus Serotype 814 strain as an Infectious Bacterial Artificial Chromosome<br>鸡马立克氏病病毒814株细菌人工染色体的构建
%A Hongyu Cui
%A Yunfeng Wang
%A Xingming Shi
%A Guangzhi Tong
%A Desong Lan
%A Lai He
%A Huaji Qiu
%A Changjun Liu
%A Mei Wang
%A <br>崔红玉
%A 王云峰
%A 石星明
%A 童光志
%A 兰德松
%A 何来
%A 仇华吉
%A 刘长军
%A 王玫
%J 生物工程学报
%D 2008
%I 
%X The aim of this study was to construct the complete genome of Marek's disease virus serotype 814 strain as an infectious bacterial artificial chromosome (BAC). Using self-designed selection marker Eco-gpt (1.3 kb) and BAC vector pBeloBAC11 (7.5 kb), we constructed the transfer plasmid pUAB-gpt-BAC11. The plasmid pUAB-gpt-BAC11 and MDV total-DNA were cotransfected into secondary CEFs; we put the virus-containing cells in selection medium for eight rounds and obtained purified recombinant viruses. Recombinant viral genomes were extracted and electroporated into E. coli, BAC clones were identified by restriction enzyme digestion and PCR analysis. Finally, we obtained 38 BAC clones, DNA from various MDV-1 BACs was transfected into CEFs, and recombinant virus was reconstituted by transfection of MDV-BAC2 DNA. We successfully cloned the complete genome of MDV-1814 strain as an infectious bacterial artificial chromosome. With these cloned genomes, a revolutionary MDV-DNA engineering platform utilizing RED/ET recombination system was constructed successfully, which can help the understanding of MDV gene functions and promote the using of MDV as a vector for expressing foreign genes. In addition, it opens the possibility to generate novel MDV-1 vaccines based on the BACs.
%K 鸡马立克氏病病毒
%K 细菌人工染色体
%K 分子克隆化病毒
%K 感染性分子克隆
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=F8845D0AA8D575ACCBEFCEEB06C04F3E&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=E158A972A605785F&sid=1FA4E9C3E6E88FC8&eid=E406B4E9A1BA9D8C&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=28