本文關(guān)鍵詞：幽門螺桿菌Ⅳ型分泌系統(tǒng)cagQ基因缺失株的構(gòu)建與鑒定　出處：《江蘇大學(xué)學(xué)報(醫(yī)學(xué)版)》2014年06期 　論文類型：期刊論文

  更多相關(guān)文章： 幽門螺桿菌 Ⅳ型分泌系統(tǒng) cag Q 基因缺失株

【摘要】：目的:構(gòu)建幽門螺桿菌(Helicobacter pylori,H.pylori)Ⅳ型分泌系統(tǒng)cag Q基因缺失株。方法:通過PCR擴(kuò)增出cag Q基因編碼區(qū)側(cè)翼同源臂序列,經(jīng)TA克隆后進(jìn)行雙酶切,并將純化后上下游同源臂連接至兩端含有卡那霉素抗生素基因的載體p Bluscript SK II(-),構(gòu)建為cag Q基因自殺質(zhì)粒。利用電穿孔法將自殺質(zhì)粒導(dǎo)入H.pylori,進(jìn)行同源重組。培養(yǎng)后通過卡那霉素抗生素篩選出基因缺失株,并經(jīng)PCR及核酸序列分析進(jìn)一步確定基因缺失株。結(jié)果:H.pylori的cag Q基因自殺質(zhì)粒p Blue KM40-△cag Q經(jīng)酶切驗(yàn)證無誤,進(jìn)行電轉(zhuǎn)化后經(jīng)PCR及核酸序列分析結(jié)果正確。結(jié)論:成功獲得H.pylori cag Q基因缺失株,命名為Hp26695-△cag Q。
[Abstract]:Objective: to construct Helicobacter pylori pylori. Methods: the flanking arm sequence of the coding region of cag Q gene was amplified by PCR and digested by double enzyme after TA cloning. After purification, the upstream and downstream homologous arms were ligated to the vector p Bluscript SK II- containing kanamycin antibiotic gene at both ends. The suicide plasmid of cag Q gene was constructed. The suicide plasmid was introduced into H. pylori by electroporation and homologous recombination. After culturing, kanamycin antibiotic was used to screen out the gene deletion strain. Results the cag Q gene suicide plasmid p Blue KM40-cag of H. pylori was further identified by PCR and nucleic acid sequence analysis. Q was confirmed by enzyme digestion. The results of PCR and nucleic acid sequence analysis were correct. Conclusion: H.pylori cag Q gene deletion strain was successfully obtained and named Hp26695- cag Q.
【作者單位】： 江蘇大學(xué)醫(yī)學(xué)院;
【基金】：江蘇省自然科學(xué)基金青年基金資助項(xiàng)目(BK20130542) 江蘇省科技創(chuàng)新與成果轉(zhuǎn)化(生命健康科技)專項(xiàng)基金資助項(xiàng)目(BL2012047) 高等學(xué)校博士學(xué)科點(diǎn)專項(xiàng)科研基金資助項(xiàng)目(20123227110008)
【分類號】：R378
【正文快照】： 幽門螺桿菌(Helicobacter pylori,H.pylori)是一類微需氧、革蘭染色陰性螺桿菌,主要定植于人胃黏膜表面及十二指腸球部[1],全球感染率達(dá)50%,慢性感染常導(dǎo)致胃炎,甚至胃十二指腸潰瘍、胃癌[2]。H.pylori中含有長度約40 kb的基因片段,即cag致病島(cytotoxin-associated gene-pat

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 梁坤;張紅珠;周廣璽;孫向紅;王輝明;張翠萍;;逆萎康對慢性萎縮性胃炎大鼠胃黏膜組織P16和TGF-β1表達(dá)影響[J];青島大學(xué)醫(yī)學(xué)院學(xué)報;2014年01期

2 陳娜;莊園;王婷婷;彭六生;程平;李文華;趙永亮;余佩武;鄒全明;;肥大細(xì)胞在胃癌患者中分布及其免疫抑制作用的研究[J];免疫學(xué)雜志;2014年01期

3 劉偉;于晗;李玉瑭;劉旭靜;李雯;單玉群;孫允東;周亞濱;;二烯丙基三硫醚對幽門螺桿菌生物膜的殺菌作用[J];山東大學(xué)學(xué)報(醫(yī)學(xué)版);2013年10期

4 謝鐵松;雷如軍;邵義敏;楊靖;劉益國;;幽門螺桿菌在胃癌形成中的作用機(jī)制[J];四川生理科學(xué)雜志;2014年01期

5 劉俊;陳德玉;趙建忠;王明義;邵世和;戴東方;;幽門螺桿菌Ⅳ型分泌系統(tǒng)Cag3基因的克隆表達(dá)及其質(zhì)譜鑒定[J];江蘇醫(yī)藥;2013年03期

6 Guo-Qing Wang;Jian-Ting Xu;Guang-Yu Xu;Yang Zhang;Fan Li;Jian Suo;;Predicting a novel pathogenicity island in Helicobacter pylori by genomic barcoding[J];World Journal of Gastroenterology;2013年30期

7 凌峰;王曉春;陳s，

本文編號：1396794