Ikeda H, Laigret F, Martin M A, Repaske R
J Virol. 1985 Sep;55(3):768-77. doi: 10.1128/JVI.55.3.768-777.1985.
The murine leukemia virus (MuLV) sequence associated with the resistance allele of the Fv-4 gene (Fv-4r) was molecularly cloned from genomic DNA of uninfected mice carrying this allele. The 5.2-kilobase cloned EcoRI DNA fragment (pFv4) was shown by nucleotide sequencing to contain 3.4 kilobases of a colinear MuLV-related proviral sequence which began in the C-terminal end of the pol region and extended through the env region and the 3' long terminal repeat. Cellular sequences flanked the 3' as well as the 5' ends of the truncated MuLV sequence. Alignment of the N-terminal half of the pFv4 env sequence with ecotropic, mink cell focus-forming, and xenotropic MuLV env sequences established the relatedness of pFv4 and ecotropic MuLV env sequences. A subcloned 700-base pair segment (pFv4env) from the 5' env region of pFv4 was used as an Fv-4-specific probe; it hybridized specifically to the Fv-4r-associated proviral sequence but not to endogenous ecotropic MuLV proviral DNA under high stringency. All Fv-4-resistant mice contained the same retroviral segment associated with the same flanking cellular DNA. Expression of Fv-4r-specific mRNA was demonstrated in the spleens of Fv-4r mice but not Fv-4s mice, supporting the previously proposed resistance model based on interference.
从小鼠基因组DNA中分子克隆了与Fv - 4基因抗性等位基因（Fv - 4r）相关的鼠白血病病毒（MuLV）序列，这些小鼠未感染病毒但携带该等位基因。通过核苷酸测序表明，5.2千碱基的克隆EcoRI DNA片段（pFv4）含有3.4千碱基的共线性MuLV相关前病毒序列，该序列始于pol区域的C末端，延伸穿过env区域和3'长末端重复序列。细胞序列位于截短的MuLV序列的3'端和5'端两侧。将pFv4 env序列的N端一半与亲嗜性、貂细胞集落形成性和异嗜性MuLV env序列进行比对，确定了pFv4与亲嗜性MuLV env序列的相关性。从pFv4的5' env区域亚克隆的700碱基对片段（pFv4env）用作Fv - 4特异性探针；在高严格条件下，它与Fv - 4r相关的前病毒序列特异性杂交，但不与内源性亲嗜性MuLV前病毒DNA杂交。所有Fv - 4抗性小鼠都含有与相同侧翼细胞DNA相关的相同逆转录病毒片段。在Fv - 4r小鼠的脾脏中检测到Fv - 4r特异性mRNA的表达，而在Fv - 4s小鼠中未检测到，这支持了先前基于干扰提出的抗性模型。
