Physical map of the linear chromosome of Streptomyces hygroscopicus 10-22 deduced by analysis of overlapping large chromosomal deletions

Xiuhua Pang, Xiufen Zhou, Yuhui Sun, and Zixin Deng*

Journal of Bacteriology 2002, 184(7):1958-1965

Epub Date: April 2002

DOI: 10.1128/JB.184.7.1958-1965.2002


The chromosomal DNA of Streptomyces hygroscopicus 10-22, a derivative of strain 5102-6, was digested with several restriction endonucleases and analyzed by pulsed-field gel electrophoresis (PFGE). Digestions with AseI gave 11 fragments with a total length of ca. 7.36 Mb. The AseI sites were mapped by analysis of overlapping chromosomal deletions in different mutants and confirmed by Southern hybridizations using partially digested genome fragments and linking cosmidsas probes. PFGE analysis of DNA with and without proteinase K treatment, together with the hybridization results, suggested a linear organization with terminal proteins and large terminal inverted repeats. Some deletion mutantshad circular chromosomes.

‘Streptomyces nanchangensis’, a producer of the insecticidal polyether antibiotic nanchangmycin and the antiparasitic macrolide meilingmycin, contains multiple polyketide gene clusters

Yuhui Sun, Xiufen Zhou, Jun Liu, Kai Bao, Guiming Zhang, Guoquan Tu, Tobias Kieser, and Zixin Deng*

Microbiology 2002, 148(Pt 2):361-371

First published: 1 February 2002

DOI: 10.1099/00221287-148-2-361


Several independent gene clusters containing varying lengths of type I polyketide synthase genes were isolated from 'Streptomyces nanchangensis' NS3226, a producer of nanchangmycin and meilingmycin.The former is a polyether compound similar to dianemycin and the latter is a macrolide compound similar to milbemycin, which shares the same macrolide ring as avermectin but has different side groups. Clusters A-H spanned about 133, 132, 104, 174, 122, 54, 37 and 59 kb, respectively. Two systems were developed for functional analysis of the gene clusters by gene disruption or replacement. (1) Streptomyces phage ΦC31 and its derived vectors can infectand lysogenize this strain. (2) pSET152, an Escherichia coli plasmid with ΦC31 attP site, and pHZ1358, a Streptomyces-Escherichia coli shuttle cosmid vector, both carrying oriT from RP4, can be mobilized from E. coli into NS3226 by conjugation. pHZ1358 was shown to be generally useful forgenerating mutant strains by gene disruption and replacement in NS3226 as wellas in several other Streptomyces strains. A region in cluster A (~133 kb) seemed to be involved in nanchangmycin production because replacement of several DNA fragments in this region by an apramycin resistance gene [aac3(IV)] gave rise to nanchangmycin non-producing mutants.

A linear plasmid temperature-sensitive for replication in Streptomyces hygroscopicus 10-22

Xiuhua Pang, Yuhui Sun, Jun Liu, Xiufen Zhou, and Zixin Deng*

FEMS Microbiology Letters 2002, 208(1):25-28

Epub Date: 30 January 2002

DOI: 10.1111/j.1574-6968.2002.tb11055.x


Streptomyces hygroscopicus 10-22 harbors a conjugative, autonomously replicating linear plasmid pHZ6 of ca. 70 kb, which shows no obvious homology with chromosomal DNA and is temperature-sensitive for replication, being stable in the host at 28 degrees C but easily lost at 37 degrees C. On a lawn of the wild-type S. hygroscopicus 10-22 cured of pHZ6, pHZ6 elicit pocks. Temperature sensitivity seemed to be a unique property for pHZ6 among six linear plasmids tested, including the well-known linear plasmids SLP2 in Streptomyces lividans 1326 and SCP1 in Streptomyces coelicolor A3(2). The distinct identity of pHZ6 from previously identified pHZ1-pHZ5 was demonstrated by the profile of relevant plasmids in six well-defined strains originated from S. hygroscopicus 10-22.


Determination of nanchangmycin and meilingmycin by high performance liquid chromatography


Yuhui Sun, Xiufen Zhou, Guoquan Tu, and Zixin Deng*

色谱 2002, 20(1):43-45

Chinese Journal of Chromatography 2002, 20(1):43-45 (Chinese)

Published online: 30 January 2002


采用高效液相色谱法同时分离测定聚醚类抗生素南昌霉素和十六元大环内酯类抗生素梅岭霉素。色谱柱为WatersX TerraTMRP18柱(3.9mm i.d.×150 mm, 5 μm),流动相为乙腈水溶液,流速为0.8 mL/min,其梯度洗脱程序为57% (体积分数,下同)乙腈(30 min)→70%乙腈(2 min)→80%乙腈(2 min)→90%乙腈(16 min)。检测波长为234 nm,柱温25℃。该法测定的灵敏度好,加标回收率高,可用于南昌链霉菌发酵液中南昌霉素和梅岭霉素的定量测定。此法具有灵敏、准确、快速的特点。

A high performance liquid chromatographic method had been established for the separation and determination of two antibiotics produced by Streptomyces nanchangensis, polyether nanchangmycin and 16-membered macrolide meilingmycin. The latter is composed of several components .The operating conditions were Waters XT erraTM RP18 column (3.9 mm i .d .×150 mm, 5 μm) at 25℃, mobile phase linear gradient elution of acetonitrile-water with 57∶43 (volume ratio) during 0 min-30 min, 70∶30 during 30 min-32 min, 80∶20 during 32 min-34 min, 90∶10 during 34 min-50 min at a flow rate of 0.8 mL/min, and photodiode array detector at 234 nm. This method is accurate, rapid and simple, and can be used for the integrated analysis of the two different natural compounds, polyethers and macrolides.