کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8420300 | 1545893 | 2018 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Application of methylation in improving plasmid transformation into Helicobacter pylori
ترجمه فارسی عنوان
استفاده از متیلاسیون در بهبود تبدیل پلاسمید به هلیکوباکتر پیلوری
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کلمات کلیدی
متیلاسیون، پلاسمید، دگرگونی، هلیکوباکتر پیلوری،
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
چکیده انگلیسی
Helicobacter pylori is an important gastrointestinal pathogen. Its strains possess different levels of powerful restriction modification systems, which are significant barriers to genetic tools used for studying the role of functional genes in its pathogenesis. Methylating vectors in vitro was reported as an alternative to overcome this barrier in several bacteria. In this study we used two H. pylori-E. coli shuttle plasmids and several single/double-crossover homologous recombination gene-targeting plasmids, to test the role of methylation in H. pylori transformation. According to our results, transformants could be obtained only after shuttle plasmids were methylated before transformation. It is helpful in gene complementation and over-expression although at a low frequency. The frequency of gene-targeting transformation was also increased after methylation, especially for the single-crossover recombination plasmids, the transformants of which could only be obtained after methylation. For the double-crossover recombination targeting plasmids, the initial yield of transformants was 0.3-0.8â¯Ãâ¯102 CFUs per microgram plasmid DNA. With the help of methylation, the yield was increased to 0.4-1.3â¯Ãâ¯102 CFUs per microgram plasmid DNA. These results suggest that in vitro methylation can improve H. pylori transformation by different plasmids, which will benefit the pathogenic mechanism research.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 150, July 2018, Pages 18-23
Journal: Journal of Microbiological Methods - Volume 150, July 2018, Pages 18-23
نویسندگان
Huilin Zhao, Linlin Xu, Qianyu Rong, Zheng Xu, Yunfei Ding, Ying Zhang, Yulong Wu, Boqing Li, Xiaofei Ji,