反刍动物支原体的细菌接合实验方案
Bacterial Conjugation Protocol for Ruminant Mycoplasmas.
作者信息
Sagné Eveline, Citti Christine, Dordet-Frisoni Emilie
机构信息
IHAP, Université de Toulouse, INRAE, ENVT, Toulouse, France.
出版信息
Bio Protoc. 2021 Jan 20;11(2):e3893. doi: 10.21769/BioProtoc.3893.
Abstract
In , two simultaneous processes of DNA transfer have been described that require direct cell-to-cell contact and are similar to conjugation. One involves the self-transmission of an integrative conjugative element (ICE) while the second concerns the horizontal transfer of large and small fragments of chromosomal DNA. Here, we describe an optimized conjugation protocol for the horizontal transfer of ICE or chromosomal DNA carrying antibiotic resistance markers (, tetracycline, gentamicin, puromycin) from donor to recipient mycoplasma cells. Calculation of the conjugation frequencies, selection and characterization of transconjugants are detailed. This protocol has been developed with but has been successfully used for and can be adapted to other related mycoplasma species.
摘要
在[具体文献]中，已经描述了两种同时发生的DNA转移过程，这两种过程都需要细胞间直接接触，并且与接合作用相似。一种涉及整合型接合元件（ICE）的自我传递，而另一种则涉及染色体DNA大小片段的水平转移。在此，我们描述了一种优化的接合方案，用于将携带抗生素抗性标记（[具体标记]、四环素、庆大霉素、嘌呤霉素）的ICE或染色体DNA从供体支原体细胞水平转移到受体支原体细胞。详细介绍了接合频率的计算、转接合子的筛选和鉴定。该方案是针对[具体支原体物种]开发的，但已成功应用于[其他支原体物种]，并且可以适用于其他相关支原体物种。
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本文引用的文献
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