Replication-competent Moloney murine leukemia virus carrying a bacterial suppressor tRNA gene: selective cloning of proviral and flanking host sequences.
Replication-competent Moloney murine leukemia virus carrying a bacterial suppressor tRNA gene: selective cloning of proviral and flanking host sequences.
复制标题
携带细菌抑制 tRNA 基因的具有复制能力的莫洛尼鼠白血病病毒：前病毒和侧翼宿主序列的选择性克隆。
DOI：
10.1073/pnas.82.4.1141
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发表时间：
1985
影响因子：
11.1
通讯作者：
R. Jaenisch
中科院分区：
文献类型：
--
作者：
W. Reik;H. Weiher;R. Jaenisch
将一个细菌抑制基因tRNA引入Moloney小鼠白血病病毒（Mo-MuLV）原病毒基因组的长末端重复序列中，构建了一个逆转录病毒，该逆转录病毒可以很容易地克隆具有侧翼宿主序列的原病毒。一种具有复制能力的病毒，Mo-MuLV sup含有tRNA琥珀色抑制基因，在组织培养细胞中复制到高滴度，并稳定地转导细菌基因。将重组病毒微注射到妊娠中期胚胎或新生小鼠体内，可以有效地进行体内复制，并表现出与野生型Mo-MuLV相同的组织趋向性。Mo-MuLV sup中的抑制基因在细菌中起作用，并允许有效地恢复原病毒基因组。从感染细胞的DNA连接到噬菌体lambda Charon 4A臂和重组噬菌体在准宿主细胞上的选择性生长表明了这一点。所有回收的噬菌体都含有Mo-MuLV原病毒序列，并且由于噬菌体lambda的高克隆能力，侧翼的宿主DNA有1-11千碱基。该病毒将有助于研究组织培养细胞和动物中病毒与宿主的相互作用。
A bacterial suppressor tRNA gene was introduced into the long terminal repeat of the Moloney murine leukemia virus (Mo-MuLV) proviral genome to construct a retrovirus that allows easy cloning of the provirus with flanking host sequences. A replication competent virus, Mo-MuLV sup containing a tRNA amber suppressor gene, was derived that replicates to high titers in tissue culture cells and stably transduces the bacterial gene. The recombinant virus can efficiently replicate in vivo when microinjected into midgestation embryos or when injected into newborn mice and displays the same tissue tropism as wild-type Mo-MuLV. The suppressor gene in Mo-MuLV sup is functional in bacteria and allows efficient recovery of proviral genomes. This was shown by ligation of DNA from infected cells to phage lambda Charon 4A arms and selective growth of recombinant phages on su- host cells. All recovered phages contained Mo-MuLV proviral sequences and, because of the high cloning capacity of phage lambda, 1-11 kilobases of flanking host DNA. This virus should facilitate studying virus-host interactions in tissue culture cells and in animals.