Yanmei Zhang

Publications (4)9.06 Total impact

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  Available from: Dongchang Sun

  Article: [Advances in the molecular mechanism of natural bacterial transformation--a review].

  Dongchang Sun, Yanmei Zhang, Yuefeng Shi
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  ABSTRACT: Naturally transformable bacteria are able to take up DNA to acquire new genetic traits in the environment. To be naturally transformed, bacteria need to establish a physiological state, called natural competence, in which DNA uptake and processing genes are expressed. DNA uptake proteins assemble a complex to pull exogenous DNA into the cytoplasm where it can recombine with the genome DNA or establish as a plasmid. In general, DNA uptake of bacteria could be divided into two stages: DNA is transported from the milieu to the periplasm at the first stage (for Gram-negative bacteria) and is translocated across the inner membrane at the second stage. Our work and other studies revealed new plasmid DNA transformation modes in Escherichia coli. Here, we first reviewed recent advances in the molecular mechanism of natural transformation and then described the distinctive plasmid transformation mode in E. coli.
  ACTA MICROBIOLOGICA SINICA 01/2012; 52(1):6-11.
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  Available from: Dongchang Sun

  Article: RpoS regulates a novel type of plasmid DNA transfer in Escherichia coli.

  Yanmei Zhang, Chunyu Shi, Jiafei Yu, Jingjing Ren, Dongchang Sun
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  ABSTRACT: Spontaneous plasmid transformation of Escherichia coli is independent of the DNA uptake machinery for single-stranded DNA (ssDNA) entry. The one-hit kinetic pattern of plasmid transformation indicates that double-stranded DNA (dsDNA) enters E. coli cells on agar plates. However, DNA uptake and transformation regulation remain unclear in this new type of plasmid transformation. In this study, we developed our previous plasmid transformation system and induced competence at early stationary phase. Despite of inoculum size, the development of competence was determined by optical cell density. DNase I interruption experiment showed that DNA was taken up exponentially within the initial 2 minutes and most transforming DNA entered E. coli cells within 10 minutes on LB-agar plates. A half-order kinetics between recipient cells and transformants was identified when cell density was high on plates. To determine whether the stationary phase master regulator RpoS plays roles in plasmid transformation, we investigated the effects of inactivating and over-expressing its encoding gene rpoS on plasmid transformation. The inactivation of rpoS systematically reduced transformation frequency, while over-expressing rpoS increased plasmid transformation. Normally, RpoS recognizes promoters by its lysine 173 (K173). We found that the K173E mutation caused RpoS unable to promote plasmid transformation, further confirming a role of RpoS in regulating plasmid transformation. In classical transformation, DNA was transferred across membranes by DNA uptake proteins and integrated by DNA processing proteins. At stationary growth phase, RpoS regulates some genes encoding membrane/periplasmic proteins and DNA processing proteins. We quantified transcription of 22 of them and found that transcription of only 4 genes (osmC, yqjC, ygiW and ugpC) encoding membrane/periplasmic proteins showed significant differential expression when wildtype RpoS and RpoS(K173E) mutant were expressed. Further investigation showed that inactivation of any one of these genes did not significantly reduce transformation, suggesting that RpoS may regulate plasmid transformation through other/multiple target genes.
  PLoS ONE 01/2012; 7(3):e33514. · 4.09 Impact Factor
• Article: Molecular analysis shows differential expression of R-spondin1 in zebrafish (Danio rerio) gonads.

  Yanmei Zhang, Fei Li, Dongchang Sun, Jiangdong Liu, Na Liu, Qixing Yu
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  ABSTRACT: R-spondin1 (RSPO1) is a potential female-determining gene in human (Homo sapiens) and mouse (Mus musculus). Its differential expression in these mammals is correlated with signaling for sex determination. As a way of studying sex determination in fish we cloned and analyzed a RSPO1 gene in zebrafish (Danio rerio). Using real-time PCR, we observed that RSPO1 is expressed more strongly in ovaries than in testes, suggesting that RSPO1 may have a role in gonad differentiation. High RSPO1 expression was detected in some non-gonadal organs like muscle and kidneys. In situ hybridization results demonstrate that RSPO1 is expressed in premature germ cells, in oogonia and primary oocytes in ovaries and in spermatogonia and spermatocytes in testes. It is also expressed in gonad somatic cells during gonadal development: in granulosa cells and theca cells of early and late cortical-alveolar stage follicles in ovaries, and in Leydig cells in testes. This differential expression may indicate that RSPO1 has a role(s) in zebrafish gonad development and differentiation. By fusing zebrafish RSPO1 with a green fluorescent protein gene, we found that RSPO1 is located in the cytosol and Golgi apparatus but not the nucleus of fish epithelioma papulosum cyprinid (EPC) cells. These preliminary findings suggest some aspects of RSPO1 like differential expression linked to sex determination may be conserved in fish while other aspects like subcellular localization differ from the mammalian RSPO1.
  Molecular Biology Reports 03/2010; 38(1):275-82. · 2.93 Impact Factor
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  Available from: Dongchang Sun

  Article: Escherichia coli is naturally transformable in a novel transformation system.

  Dongchang Sun, Yanmei Zhang, Yunjun Mei, Hui Jiang, Zhixiong Xie, Huihui Liu, Xiangdong Chen, Ping Shen
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  ABSTRACT: A novel transformation system, in which neither a nonphysiological concentration of Ca2+ and temperature shifts nor electronic shocks were required, was developed to determine whether Escherichia coli is naturally transformable. In the new protocol, E. coli was cultured normally to the stationary phase and then cultured statically at 37 degrees C in Luria-Bertani broth. After static culture, transformation occurred in bacteria spread on Luria-Bertani plates. The protein synthesis inhibitor chloramphenicol inhibited this transformation process. The need for protein synthesis in plated bacteria suggests that the transformation of E. coli in this new system is regulated physiologically.
  FEMS Microbiology Letters 01/2007; 265(2):249-55. · 2.04 Impact Factor