一种新的编码霍乱毒素的丝状噬菌体CTAKΦ

来源 :中华微生物学和免疫学杂志 | 被引量 : 0次 | 上传用户:ruocich
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
目的 探讨介导霍乱弧菌毒素基因水平转移的一种独特的遗传结构。方法 从O139群霍乱弧菌菌株FJ9712 9上清中分离出丝状噬菌体颗粒CTAKΦ ,并进行电镜观察。从丝状噬菌体颗粒CTAKΦ中纯化出DNA ,并进行链型分析。对pCTAK进行酶谱分析。用ctxAB、zot引物对pCTAK进行PCR扩增检测 ,用RS1探针对pCTAK进行Southernblot检测 ;用pCTAK转化DH5α和IEM10 1得到DX2 9和 432 9,将pCTAK克隆于pUC18载体并转入DH5α得到UD2 9,用GM1 ELISA检测DX2 9、432 9、UD2 9的CT表达。对pCTAK的ctxAB、zot基因片段进行测序 ,并用DNASTAR软件和BLAST算法 ,在国际互联网上对测序结果进行序列分析。结果 发现和分离了一种编码霍乱毒素的质粒pCTAK ,它以稳定的高拷贝数存在于 1株天然的O139霍乱弧菌FJ9712 9中 ;酶谱分析发现其明显不同于CTX元件酶谱 ,在CTX元件中相当保守的酶切位点 :BglⅡ、EcoRⅤ、PstⅠ、EcoRⅠ ,在pCTAK中没有切点。ctxAB、zot引物对pCTAK的PCR扩增检测呈阳性 ,RS1探针杂交呈阳性 ;pCTAK的ctxAB、zot基因序列与已报道的序列有很高的同源性。pCTAK能直接或经载体转化DH5α和IEM10 1并表达CT。从FJ9712 9培养上清中分离出丝状噬菌体颗粒CTAKΦ ,电镜观察并计算其直径大小为 7nm左右。其全基因组大小为 8.5kb ,为? Objective To investigate a unique genetic structure that mediates horizontal transfer of Vibrio cholera toxin genes. Methods The filamentous phage particles CTAKΦ were isolated from the supernatant of Vibrio cholerae O139 strain FJ97129 and observed under electron microscope. DNA was purified from filamentous phage particles CTAK [Phi] and subjected to a strand analysis. Enzymatic analysis of pCTAK. PCR amplification of pCTAK was carried out using ctxAB and zot primers, and Southern blot was performed on pCTAK using RS1 probe; DH5α and IEM101 were transformed into pXD9 and pGEM9 by pCTAK; pCTAK was cloned into pUC18 vector and transformed into DH5α to obtain UD2 9 The CT1 expression of DX2 9, 432 9 and UD2 9 was detected by GM1 ELISA. The ctxAB and zot gene fragments of pCTAK were sequenced and the sequencing results were sequenced on the Internet using DNASTAR software and BLAST algorithm. Results A cholera toxin-encoding plasmid, pCTAK, was found and isolated in a stable high copy number in a native strain of V. cholerae FJ97129; zymogram analysis revealed significant differences from the CTX element zymogram and in CTX Components quite conservative sites: Bgl Ⅱ, EcoRⅤ, Pst Ⅰ, EcoR Ⅰ, no point in the pCTAK. The ctxAB and zot primers were positive for PCR amplification of pCTAK and the RS1 probe was positive for hybridization. The sequences of ctxAB and zot genes of pCTAK were highly homologous with the reported sequences. pCTAK can transform DH5α and IEM101 directly or via vectors and express CT. Filamentous phage particles CTAKΦ were isolated from the culture supernatant of FJ97129. The diameter of the filamentous phage was about 7nm by electron microscopy. The whole genome size is 8.5kb, for?
其他文献
分别用 10 %草甘磷水剂 4 0倍、60倍、80倍、10 0倍液作枳砧苗圃除草试验。结果表明 ,这几种浓度草甘磷对枳苗无伤害 ,不影响枳苗生长 ,对杂草防除效果显著 ,防效达 95 %以上
目的研究不同全麻方法纳屈酮快速脱毒的安全性和有效性.方法选择自愿接受全麻辅助下快速阿片脱毒患者160例,随机分为两组.A组(n=78),采用异丙酚、咪唑安定复合氯胺酮麻醉;B组
PCB1254对蛋白核小球藻Cholrella pyrenoidosa和斜生栅藻为Scenedesmus obliguus 2种藻类的生长有比较明显的影响,在低质量浓度时(2.0 ng/mL)时表现明显的抑制作用.2种藻类的
用SPF鸡胚从我国东北地区猪群中分离出3株H3N2猪流感病毒(SIV),分别命名为A/Swine/Heilongjiang/1/2000、A/Swine/Heilogjiang/2/2000和A/Swine/Heilogjiang/3/2000.SIV分离
To exert strong actuating power of piezoelectric stack and facilitate the incorporation of piezoelectric stack and host structure for vibration control applicat
两种有代表性的紫色土的水稻盆栽试验结果表明 ,磷的加入可减轻受砷污染的红紫泥中砷对水稻的毒害 ;但磷的加入会加剧受砷污染的红棕紫泥中砷对水稻的毒害。根际无机砷的形态
A high performance white light emitter diode (LED) driver based on boost converter with novel single-wire serial-pulse digital dimming (SWSP) is proposed. The d
以油菜品种甘杂 1号为材料 ,研究应用BN - 3、BN - 4调节剂对促进油菜生长发育的作用。结果表明 :喷施BN使油菜生育期提前 1~ 2d ,增加角果面积 2 2 .99~ 32 4 .6 0cm2 /株 ,增
Content addressable storage (CAS) is a promising technology for improving storage efficiency as well as access throughput.Currently,many CAS products are implem
本文在水分活度(aw)的蒸汽压测定法(VPM)的研究中发现Lewicki的测定装置及计算公式中尚未考虑到干燥瓶的容积和干燥剂吸湿后剩余水汽压强对aw测定结果的影响.本研究在Lewicki