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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >A broad-range of recombination cloning vectors in mycobacteria.
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A broad-range of recombination cloning vectors in mycobacteria.

机译：分枝杆菌中广泛的重组克隆载体。

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摘要

The increased incidence of tuberculosis (TB) gave impetus for the increased interest in the study of mycobacterial genetics, which culminated in the publication of the full genome sequence of many mycobacterial strains. Since then, many genes and open reading frames of unknown function have been described and the expression of their encoded proteins is critical toward understanding the pathogenesis of TB and developing therapeutic and preventive strategies. Therefore there is an increased need for highly efficient methods for cloning of mycobacterial genes, as the limited cloning flexibility of current Escherichia coli-mycobacteria shuttle vectors remains a frequent impediment in genetic manipulation of mycobacteria. In order to overcome this limitation, we have converted representative extrachromosomal and integrative vectors into multiple destination mycobacterial vectors for one-step and restriction enzyme-free recombination cloning methodology that uses in vitro site-specific recombination. We provide several examples that highlight the potential of recombination cloning for gene expression in slow and fast-growing mycobacteria. Thus, a gene of interest can be transferred by simple recombination into our mycobacterial destination vectors, which serve a multitude of functional genomic studies.
机译：结核病（TB）发病率的增加推动了对分枝杆菌遗传学研究兴趣的增长，最终导致许多分枝杆菌菌株全基因组序列的发布。从那时起，已经描述了许多功能未知的基因和开放阅读框，其编码蛋白的表达对于理解结核病的发病机理以及制定治疗和预防策略至关重要。因此，由于目前的大肠杆菌-分枝杆菌穿梭载体的有限的克隆灵活性仍然是分枝杆菌基因操作中的常见障碍，因此越来越需要高效的方法来克隆分枝杆菌基因。为了克服这一局限性，我们已将代表性的染色体外和整合型载体转化为多个目的分枝杆菌载体，以进行一步和无限制性酶的重组克隆方法，该方法使用了体外位点特异性重组。我们提供了几个示例，突出了在慢速和快速增长的分枝杆菌中重组克隆进行基因表达的潜力。因此，可以通过简单重组将目的基因转移到我们的分枝杆菌目的载体中，该载体可进行多种功能基因组研究。

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