栖热菌噬菌体TSP4密码子偏嗜性及其对基因异源表达的影响

 为了探索噬菌体TSP4、栖热菌模式菌株Thermus thermophilus HB27中6种蛋白编码序列和Escherichia coli基因组遗传密码子使用偏嗜性,探索TSP4基因密码子偏嗜性对其基因异源表达的影响本研究利用生物学软件Editseq和RSCU算法统计噬菌体TSP4密码子使用频率并分析其偏嗜性,与栖热菌HB27的同源基因以及常温菌E.coli基因组的密码子使用偏嗜性进行对比分析.结果显示,噬菌体TSP4与栖热菌HB27优势密码子相似度高达85%,而与E.coli的相似性仅有65%.为了验证分析结果,选取TSP4基因组中一个潜在分子伴侣基因序列在E.coli BL21和E.coli Rosetta(补充了BL21菌株中缺失的6个稀有密码子)中进行异源表达.噬菌体TSP4与其宿主菌HB27之间密码子高相似性说明在长期的进化过程中TSP4在基因水平上形成对宿主的一种适应性机制.异源表达结果显示,分子伴侣基因在E.coli BL21中未见明显表达,但在Rosetta中高效表达,说明Rosetta中补充的 6个稀有密码子明显有助于分子伴侣基因的表达,该结果也进一步证明密码子偏嗜性是否一致在很大程度上影响基因的表达.     

滇西二高温温泉中Thermus高温菌的16S rRNA

 通过对滇西二温泉纯培养和免培养法获得的Thermus菌株序列进行16SrDNA系统发育及16SrRNA二级结构比较,发现二温泉中至少存在Thermus属的2个潜在新种,它们在系统发育树上形成1个独立的地理簇群分支,它们的16SrRNAhelix10二级结构也与其它地区的Thermus属菌株不同,形成明显的地域进化特征.     

Cesium accumulation by bacterium Thermus sp. TibetanG7： hints for biomineralization of cesiumbearing geyserite in hot sprinss in Tibet, China

The bacterium Thermus sp. TibetanG7, isolated from hot springs in Tibet, China, was examined for the ability to accumulate cesium from solutions. Environmental conditions were simulated and the effects of pH, K^＋, Na^＋and K^＋-regimes were then studied to determine the possible role of the bacterium in the formation of cesium-bearing geyserite around these hot springs. In despite of the inhibition of K^＋ and Na^＋, the bacterium Thermus sp. TibetanG7 revealed noticeable accumulation of cesium from solutions, with maximum accumulations of 53.49 and 40.41 pmol Cesium/g cell dry weight in Na^＋ and K^＋ inhibition experiments, respectively. The accumulation of cesium by this microorganism is rapid, with 40% --50% accumulated within the first 5 min. K^＋-deficient cells showed a much higher capacity of cesium accumulation compared with K^＋-sufficient cells. It is evident that the bacteria within the genus thermus play a significant role in the cesium assembly. The formation of cesium-bearing geyserite is also considered.     

大理洱源地区高氟热泉中栖热菌属细菌(Thermus sp)的研究初报

从洱源地区高氟热泉中分离得到３６株专性嗜热非芽孢杆菌,经鉴定为栖热菌属（Ｔｈｅｒｍｕｓ）,根据生理生化特性测定,３６株菌又分为两个类群：ＰＡ群２２株菌氧化酶阳性、不产色素,ＰＢ群１４株菌氧化酶阴性、产黄色色素．同时对３６株菌的分布作了一些初步探讨．     

Cesium accumulation by bacterium Thermus sp. TibetanG7:hints for biomineralization of cesium-bearing geyserite in hot springs in Tibet,China

The bacterium Thermus sp. TibetanG7, isolated from hot springs in Tibet, China, was examined for the ability to accumulate cesium from solutions. Environmental conditions were simulated and the effects of pH, K , Na and K -regimes were then studied to determine the possible role of the bacterium in the formation of cesium-bearing geyserite around these hot springs. In despite of the inhibition of K and Na , the bacterium Thermus sp. TibetanG7 revealed noticeable accumulation of cesium from solutions, with maximum accumulations of 53.49 and 40.41 μmol Cesium/g cell dry weight in Na and K inhibition experiments, respectively. The accumulation of cesium by this microorganism is rapid, with 40%―50% accumulated within the first 5 min. K -deficient cells showed a much higher capacity of cesium accu- mulation compared with K -sufficient cells. It is evident that the bacteria within the genus thermus play a significant role in the cesium assembly. The formation of cesium-bearing geyserite is also considered.     

云南热泉栖热菌属细菌(Thermussp)的质粒研究

从云南汤池、腾冲热泉（ｐＨ７．０～９．０，７０～９２℃）分离到８株专性嗜热非芽孢杆菌，最适生长温度６５～７５℃．鉴定为栖热菌属（Ｔｈｅｒｍｕｓ），根据生理生化特性测定，８株菌又分为两个群，分离自汤池样点的ＴＡ群５株菌氧化酶阳性，分离自腾冲样点的ＴＢ群３株菌，氧化酶阴性．通过质粒快速抽提，琼脂糖凝腋电脉发现，８株菌都含有一个质粒，大小略大于８．５ｋｂ．     

栖热菌(Thermus)的DNA转化研究

由栖热菌FDB8(Thermus sp.FDB8)提取的染色体DNA对栖热菌FD3009(Thermus sp.FD3009)进行了转化.用丝裂霉素C选择培养基对转化子进行选择,共获得3个转化子.它们的菌落色泽、对丝裂霉素C的抗性强度和耐热DNA聚合酶活性均介于供体菌和受体菌之间,而其菌体生长速度和破壁难易程度则优于供体菌和受体菌.结果表明,转化子是由供体菌FDB8的染色体DNA转化了受体菌FD3009所致,同时还可由转化子进一步选育出抗丝裂霉素C的耐热DNA聚合酶高产菌株.     

嗜热菌Thermus thermophilus HB8铁硫簇合成途径中IscA和IscU蛋白的表达、纯化和功能研究

Fe-S簇可参与电子传递和基因调节等过程.实验以嗜热菌Thermus thermophilus HB8基因组为模板,通过PCR扩增获得Fe-S簇ISC系统生物合成途径中的IscA和IscU基因,并克隆连接到表达载体pET-28a（＋）上.重组质粒转化E.coli Rosetta（DE3）进行表达,用亲和层析纯化得到浓度和纯度都较高的IscA和IscU蛋白.SDS-PAGE分析,证实其分子量大小分别为13.7KD和14.8KD.实验确定IscA是单聚体,IscA上能形成铁硫簇,IscU上不能形成铁硫簇.在厌氧条件下,RNA的量能提高1.5倍,但是IscA和IscU的表达量都会降低,说明厌氧对这两个基因有损伤.