质粒RSF1010从大肠杆菌到稀有放线菌头状链轮丝菌和星状诺卡氏菌的接合转移

１９８７年,ＴｒｉｅｎＣｕｏｔ等人［１］证明穿梭质粒可以在革兰氏阴性的大肠杆菌（Ｅｓｃｈｅｒｉｃｈｉａｃｏｌｉ）和多种革兰氏阳性细菌之间发生接合转移。在这种转移中质粒需具备大肠杆菌的复制起始位点,同时又具备革兰氏阳性细菌的广宿主范围复制起始位点。转…     

诺卡氏菌型放线菌的分类

诺卡氏菌型放线菌在分类学上属于细菌域 ,厚壁菌门 ,放线细菌纲 ,放线菌目中的一类形态相似的微生物。在自然界中分布广泛 ,但由于分离、分类方法所限 ,绝大多数 ( 85%以上 )还没有被人们所认识。诺卡氏菌型放线菌是一类重要的自然资源 ,某些种能产生抗生素 ,如利福毒素等 ;有些种产生可供利用的重要的代谢产物 ,如 2、3 二羟基 苯甲酸、环已乙酸、羟基苯酮丁酸、Ｌ 组氨酸、1 6α 羟基甾体等 ;某些种产生重要的酶类 ,如胆固醇氧化酶、葡萄糖异构酶、溶菌酶等 ;有些种可用于废水 (物 )的生物处理。另一方面 ,也有些种引起人和动物的诺卡氏…     

诺卡氏菌形放线菌枝菌酸的气相色谱分析

以已知的棒杆菌枝菌酸,红球菌枝菌酸及自诺卡氏菌典型株提取的枝菌酸为参照样品,用硅烷化的枝菌酸甲酮气相色谱分析方法,分析侧定了包括以前我们发表的两个诺卡氏菌种在内的7株诺卡氏菌形放线菌的枝菌酸。结果表明.鲜黄诺卡氏菌A100和黄粉诺卡氏菌N10的枝菌酸分子碳原子数(58-64)均在诺卡氏菌枝菌酸的范围内;而另一株原名珊瑚诺卡氏菌N21的枝菌酸碳原子数(32-40)则与红色红球菌8372一样在红球菌枝菌酸范围内。其余结果均与文献报道的相一致。本文还讨论了这一方法在诺卡氏菌形放线菌分类鉴定中的作用和意义。     

诺卡氏菌形放线菌的化学分类

通过对28株分别代表12个属的诺卡氏菌形放线菌及有关分类单位的胞壁组分、枝菌酸、甲基萘醒、磷酸类脂等化学特性的分析,结合数值分类,在相似值(s.m)80％水平上将这类菌分成了两个大的化学类群,在90％水平上再分成四个小的类群。除拟诺卡氏菌属是显著的化学异源类群外,其余均与传统分类的属一致,形成独立的化学分类单位。讨论了化学分类在诺卡氏菌形放线菌分类研究中的应用及对出现的化学异源类群的认识。     

拟诺卡氏菌线型质粒pNPL1的测序和分析

【目的】检测和分析稀有放线菌中新的线型质粒。【方法】从植物内生菌中分离链霉菌之外的放线菌菌株,检测、测序和分析线型质粒。【结果】从中草药植物紫花前胡的叶片中分离到一株内生放线菌25L-1-1c,经过16S rRNA基因序列比对属于拟诺卡氏菌。从该菌株中检测到一个约25 kb的线型质粒pNPL1。克隆和测序了pNPL1新的端粒,含有多个小的回文序列。测序获得全长为24 621 bp的线型质粒pNPL1,预测编码22个基因,其中2个基因与链霉菌质粒的端粒复制基因同源,1个基因与链霉菌质粒主要的接合转移基因相似,其余19个基因为未知功能。携带pNPL1端粒复制基因的质粒不能转化变铅青链霉菌,暗示需要发展拟诺卡氏菌的遗传操作系统。【结论】这是首次在拟诺卡氏菌中发现和描述线型质粒。     

诺卡氏菌形放线菌β-甘露聚糖酶的纯化和性质

产β－１,４－Ｄ甘露聚糖酶的诺卡氏菌形放线菌（Ｎｏｃａｒｄｉｏｆｏｒｍ ａｃｔｉｎｏｍｙｃｅｔｅｓ）菌株ＮＡ３－５４０,发酵培养７２ｈ,发酵液离心去菌体,上清经硫酸铵沉淀,９５％乙醇沉淀,ＣＭ－Ｓｅｐｈａｄｅｘ Ａ５０柱层析、羟基磷灰石柱层析、ＤＥＡＥ－纤维素离子交换及Ｓｅｐｈａｄｅｘ Ｇ－１００分子凝胶过滤柱等步骤,β－甘露聚糖酶的比活提高了１３７倍,获得凝胶电泳均一的蛋白样品。经ＳＤＳ－ＰＡＧ     

诺卡氏菌形放线菌与有关分类单位的枝菌酸和磷酸类脂分析

通过对14株胞壁为I型和IV型的诺卡氏菌形放线菌与有关分类单盘的枝菌酸和磷酸类脂的分析,发现胞壁IV型原名为空腔诺卡氏菌N. coeliaca KCC A-0007菌株不含枝菌酸,磷酸类脂不属于任何已知类型(除含PE外,还含有PC),萘醌类型为MK-8(H4)。指出这株菌应从诺卡氏菌属转入无枝菌酸菌属,改定为空腔无枝菌酸菌Amycolata coeliaca[(Gray)Waksrnanet Henrici] comb. Nov. Liu ＆ Ruan,1986。另外还提出,区分类诺卡氏菌与链霉菌时,磷酸类脂类型比形态断裂特征更为重要。为此,将一株原定名为阿勒泰类诺卡氏菌Nocardioides al-taiensis (Wang,1982),依其磷酸类脂分析结果,与链霉菌(Ⅱ型)相同,而与类诺卡氏菌(I型)不同,改定为阿勒泰链霉菌Streptomyces altaiensis (Wang) comb．Nov．Liu ＆ Ruan, 1986。     

诺卡氏菌型放线菌细胞中脂肪酸的气相色谱分析

采用Sherolock全自动微生物鉴定系统,用气相色谱法,分析测定了4株诺卡氏菌型放线菌分离菌株的脂肪酸成分和含量,结果表明,该法具有分辨率高,稳定、重现性好,简便易行等特点,在一定程度上与16S rRNA基因序列比较结果相一致,能在种及菌株水平上反映出放线菌的基因型,系统发育和分类关系,是一种较好的脂肪酸定量测定方法,尤其适用于分析大量的菌株或分离株,可应用于放线菌种水平的分类和快速鉴定.     

诺卡氏菌科分类的研究II．诺卡氏菌属中的两个新种和一个新变种

从我国各地的土壤中分离得到百余株诺卡氏菌形放线菌,其基丝形成横隔并断裂成秆状或球状体,细胞壁化学组份IV型,属于诺卡氏菌科诺卡氏菌属(Nocardia)。经形态、培养特征及生理生化特性等鉴定为15个种。本文只报道其中两个新种及一个新变种：念球状诺卡氏菌(Noeaedia nostocoidea n. sp．);紫褐诺卡氏菌(Nocardta violaccofusca n. sp.);鲑色诺卡氏菌桔橙变种(Nocardia salmontcolor var. aurantiaca n. var.)。     

Molecular cloning of the structural genes of the Escherichia coli deo-operon in plasmid RSF2124]

A specialized phage lambda ddeo carrying the deo operon of Escherichia coli is analyzed by exposing the DNA to the specific restriction endonucleases EcoRI and BamHI. Using the lambda ddeo DNA fragment, obtained by digestion with BamHI and plasmid RSF2124 as a vehicle, the hybrid plasmid pAM1 carrying all the genes of the deo operon is constructed and cloned in E. coli cells. It is shown that the activity of thymidine phosphorylase in the strain AM061, which contains hybrid plasmid pAM1 is 30-fold greater than that in strains of E. coli with chromosomal localization of the deo operon.     

Transfer of plasmid RSF1010 by conjugation from Escherichia coli to Streptomyces lividans and Mycobacterium smegmatis.

The plasmid RSF1010 belongs to a class of plasmids (IncQ) that replicate in a range of bacterial hosts. Although non-self-transmissible, it can be mobilized at high frequency between different gram-negative bacterial species if transfer functions are supplied in trans. We report the transfer of RSF1010 by conjugation from Escherichia coli to the gram-positive actinomycetes Streptomyces lividans and Mycobacterium smegmatis. In its new hosts, the plasmid was stable with respect to structure and inheritance and conferred high-level resistance to streptomycin and sulfonamide. This is the first reported case of conjugative transfer of a naturally occurring plasmid between gram-negative and gram-positive bacteria.     

Replication of the nonconjugative plasmid RSF1010 in Escherichia coli K-12.

Replicating DNA molecules of the nonconjugative R plasmid RSF1010 (Smr Sur) were cleaved with the EcoRI restriction endonuclease and examined with the electron microscope. Results of this analysis indicated that replication is initiated from an origin located at about 19% of total genome size from one of the EcoRI ends. Replication proceeded either unidirectionally or bidirectionally with equal frequency. Results of the analysis of replicative intermediates of RSF1010 containing the Apr-transposable sequence (Tn) are also presented.     

Maintenance of plasmid pSC101 in Escherichia coli requires the host primase.

The abilities of three Escherichia coli strains with thermosensitive dnaG alleles to maintain plasmids pSC101 or pBR322 or an RP4 derivative were studied at elevated growth temperatures. Under these conditions, pSC101 segregated from cells to a greater extent than did pBR322. No segregation of the primase-encoding RP4 derivative was observed.     

Transmission of F'lac plasmid from Escherichia coli K-12 to bacteria of the genus Erwinia]

The F'lac plasmid was transferred by conjugation from Escherichia coli K-12 W1655 to 21 lac- strains of Erwinia spp. (5.2 . 10(-6) to 6.8 . 10(-2) lac+ exconjugants per donor cell). Erw. herbicola and Erw. chrysanthemi were the better recipients than others. The degree of the stability of lac+ genes in Erwinia exconjugants depends on the strains. Stable exconjugants of Erwinia, which harbored F'lac plasmid, were able to utilize lactose, to transfer lac genes by conjugation to Erwinia spp. and E. coli, and were sensitive to the F-specific phages f1, f2, Qbeta. The F'lac plasmid was eliminated from the exconjugants by the treatment with acridine orange, which indicates that this genetic element is not integrated into the Erwinia chromosome.     

A plasmid carrying mucA and mucB genes from pKM101 in Haemophilus influenzae and Escherichia coli.

The plasmid pMucAMucB, constructed from the Haemophilus influenzae vector pDM2, and a similar plasmid, constructed from pBR322, increased the survival after UV irradiation of Escherichia coli AB1157 with the umu-36 mutation and also caused UV-induced mutation in the E. coli strain. In H. influenzae, pMucAMucB caused a small but reproducible increase in survival after UV irradiation in wild-type cells and in a rec-1 mutant, but there was no increase in spontaneous mutation in the wild type or in the rec-1 mutant and no UV-induced mutation.     

TnA-directed deletion of the trp operon from RSF2124-trp in Escherichia coli

Plasmid pMT-trp was constructed by digestion of RSF2124-trp with restriction endonuclease PstI and ligation with T4 ligase. In pMT-trp about 78% of the DNA of transposon TnA from RSF2124-trp was deleted, and hence the gene for ampicillin resistance was lost. All Trp- segregants from pMT-trp carriers in Escherichia coli W3110 and its derivatives were found to have lost the entire plasmid. On the other hand, deletion plasmids which had lost the trp operon were found among Trp- segregants from RSF2124-trp carriers, particularly from the mutant strain trpAE1 trpR tnaA. The experimental fact that deletion occurred exclusively in RSF2124-trp suggests that the presence of TnA in the plasmid (RSF2124-trp) was responsible for the deletion.