Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12.

J D van Embden; H J van der Donk; R V van Eijk; H G van der Heide; J A de Jong; M F van Olderen; A B Osterhaus; L M Schouls

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Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12.

机译：大肠杆菌K-12中蛋白酶Pallidum DNA的分子克隆和表达。

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A gene bank of Treponema pallidum DNA in Escherichia coli K-12 was constructed by cloning SauI-cleaved T. pallidum DNA into the cosmid pHC79. Sixteen of 800 clones investigated produced one or more antigens that reacted with antibodies from syphilitic patients. According to the separation pattern of the antigens produced on sodium dodecyl sulfate-polyacrylamide gels, six different phenotypes were distinguished among these 16 clones. These antigens reacted also with anti-T. pallidum rabbit serum. No antibodies against the cloned antigens were found in normal rabbit serum and in nonsyphilitic human serum. The antigens produced by the E. coli K-12 recombinant DNA clones comigrated in sodium dodecyl sulfate-polyacrylamide gels with antigens extracted from T. pallidum bacteria, suggesting that the treponemal DNA is well expressed in E. coli K-12. Several of the cosmid recombinant plasmids have been subcloned, resulting in smaller T. pallidum recombinant plasmids which are more stably maintained in the cell and produce more treponemal antigen. Monoclonal antibodies were raised against T. pallidum, and one hybridoma produced antibodies that reacted not only with an antigen from T. pallidum but also with the antigen produced by one of the E. coli clones.

机译：通过将Saui-Cleaved T.Pallidum DNA克隆到粘性PHC79中构建了大肠杆菌C大肠杆菌K-12中的蛋白酶粘液蛋白DNA的基因库。 800个克隆的克隆研究产生了一种或多种与来自梅氏患者的抗体反应的抗原。根据在十二烷基硫酸钠 - 聚丙烯酰胺凝胶上产生的抗原的分离模式，在这16个克隆中区分了六种不同的表型。这些抗原也与抗T反应。 Pallidum兔血清。在正常的兔血清和非尿嘧啶人血清中没有发现抗克隆抗原的抗体。由大肠杆菌K-12重组DNA克隆生产的抗原在二烷基硫酸钠 - 聚丙烯酰胺凝胶中，用从T.Pallidum细菌提取的抗原，表明纤维型DNA在大肠杆菌K-12中良好地表达。已经亚克隆了几种粘性重组质粒，导致缺点粘性重组质粒，其更稳定地保持在细胞中并产生更多的抗精氨酸抗原。单克隆抗体升高针对磷酸盐，并且一种杂交瘤产生的抗体不仅与来自T.Pallidum的抗原反应，而且与由大肠杆菌克隆之一产生的抗原反应。

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《Infection and immunity》 |1983年第1期|共10页
作者
J D van Embden; H J van der Donk; R V van Eijk; H G van der Heide; J A de Jong; M F van Olderen; A B Osterhaus; L M Schouls;
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1. Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12. [J] . J D van Embden, H J van der Donk, R V van Eijk, Infection and immunity . 1983,第1期

机译：大肠杆菌K-12中蛋白酶Pallidum DNA的分子克隆和表达。
2. Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12. [J] . J D van Embden, H J van der Donk, R V van Eijk, Infection and immunity . 1983,第1期

机译：大肠杆菌K-12中蛋白酶Pallidum DNA的分子克隆和表达。
3. Expression of Treponema pallidum antigens in Escherichia coli K-12. [J] . L V Stamm, J D Folds, P J Bassford Infection and immunity . 1982,第3期

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4. Isolation, Molecular Cloning and Expression of Cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli [C] . Woon J.S.K., Murad A.M.A., Abu Bakar F.D. UKM FST Postgraduate Colloquium . 2015

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6. Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12. [O] . J D van Embden, H J van der Donk, R V van Eijk, 1983

机译：梅毒螺旋体DNA的分子克隆和表达在大肠杆菌K-12中。
7. Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K12. [O] . Embden J.D.A. van, Donk H.J.M. van de, Eijk R.V.W. (Ron) van, 1983

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8. Cloning and Expression in Escherichia coli of Clostridium thermocellum DNAEncoding Subcellulosomal Proteins [R] . Kobayashi, T., Romaniec, M. P., Fauth, U., 1992

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Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12.

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