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[用特异性抗体筛选日本血吸虫成虫基因组DNA文库]

[Screening of Schistosoma japonicum adult worm genomic DNA library by specific antibody].

作者信息

Tao Y W, Xu Y X, Liu S X, Zheng Z X, Yan W Y

机构信息

Institute of Parasitic Disease, Chinese Academy of Preventive Medicine.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1990;8(4):264-6.

PMID:2099255
Abstract

S. japonicum adult worm genomic DNA libraries were screened by enzymeimmunoassay. The antigens produced by recombinant lambda gt 11 plaques were transferred to nitrocellulose filters. Clones encoding given antigens were detected with infected rabbit sera (IRS) which were preabsorbed with lysate of induced lambda gt 11 in Y1090 cells. Eight putative positives were picked up from 97 plates in the primary screening and were rescreened a second time, and, 2 of them consistently gave good positive signals in subsequent screenings. One of the phage clones was purified to homogeneity and mixed with Y1089 cells. The expressed products still showed positive when rescreened by ELISA, indicating that the clone encoding S. japonicum antigen could be recognized by IRS. The immunoscreening method described here was able to efficiently isolate single clones encoding S. japonicum antigens. The method has the advantages of screening libraries efficiently, reliably and specifically, and it might open the way to screen genes encoding S. japonicum antigens inducing protective immunity (Fig. 1).

摘要

用酶免疫测定法筛选日本血吸虫成虫基因组DNA文库。将重组λgt 11噬菌斑产生的抗原转移至硝酸纤维素滤膜上。用经Y1090细胞中诱导型λgt 11裂解物预吸附的感染兔血清(IRS)检测编码特定抗原的克隆。在初次筛选中,从97个平板中挑选出8个假定阳性克隆并进行二次筛选,其中2个在后续筛选中始终给出良好的阳性信号。纯化其中一个噬菌体克隆至同质性,并与Y1089细胞混合。通过ELISA重新筛选时,表达产物仍呈阳性,表明编码日本血吸虫抗原的克隆可被IRS识别。本文所述的免疫筛选方法能够有效分离编码日本血吸虫抗原的单一克隆。该方法具有高效、可靠且特异的文库筛选优势,可能为筛选编码诱导保护性免疫的日本血吸虫抗原的基因开辟道路(图1)。

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