Cordingley J S, Taylor D W, Dunne D W, Butterworth A E
Gene. 1983 Dec;26(1):25-39. doi: 10.1016/0378-1119(83)90033-1.
We have constructed a small vector specifically for blunt-end cloning of fragments of DNA. Both the PvuII site and the EcoRI site allow the detection of recombinants using a simple and inexpensive colour screen. We have used this vector to construct cDNA clone banks from polyadenylated messenger RNA [poly(A)+mRNA] from several life cycle stages of the human parasite Schistosoma mansoni and have identified clones encoding an immunodiagnostic antigen gene by a combination of Southern blotting and mRNA hybrid-selection and in vitro translation. Antibodies against this antigen are only present in patients infected with S. mansoni.
我们构建了一种专门用于DNA片段平端克隆的小载体。PvuII位点和EcoRI位点都能通过简单且成本低廉的颜色筛选来检测重组体。我们已使用该载体从人类寄生虫曼氏血吸虫几个生命周期阶段的聚腺苷酸化信使RNA[poly(A)+mRNA]构建了cDNA克隆文库，并通过Southern印迹、mRNA杂交选择和体外翻译相结合的方法鉴定出编码免疫诊断抗原基因的克隆。针对这种抗原的抗体仅存在于感染曼氏血吸虫的患者体内。
