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高频转化海洋弧菌菌株中的天然质粒转化

Natural plasmid transformation in a high-frequency-of-transformation marine Vibrio strain.

作者信息

Frischer M E, Thurmond J M, Paul J H

机构信息

Department of Marine Science, University of South Florida, St. Petersburg 33701-5016.

出版信息

Appl Environ Microbiol. 1990 Nov;56(11):3439-44. doi: 10.1128/aem.56.11.3439-3444.1990.

Abstract

The estuarine bacterium Vibrio strain DI-9 has been shown to be naturally transformable with both broad host range plasmid multimers and homologous chromosomal DNA at average frequencies of 3.5 X 10(-9) and 3.4 X 10(-7) transformants per recipient, respectively. Growth of plasmid transformants in nonselective medium resulted in cured strains that transformed 6 to 42, 857 times more frequently than the parental strain, depending on the type of transforming DNA. These high-frequency-of-transformation (HfT) strains were transformed at frequencies ranging from 1.1 X 10(-8) to 1.3 X 10(-4) transformants per recipient with plasmid DNA and at an average frequency of 8.3 X 10(-5) transformants per recipient with homologous chromosomal DNA. The highest transformation frequencies were observed by using multimers of an R1162 derivative carrying the transposon Tn5 (pQSR50). Probing of total DNA preparations from one of the cured strains demonstrated that no plasmid DNA remained in the cured strains which may have provided homology to the transforming DNA. All transformants and cured strains could be differentiated from the parental strains by colony morphology. DNA binding studies indicated that late-log-phase HfT strains bound [3H]bacteriophage lambda DNA 2.1 times more rapidly than the parental strain. These results suggest that the original plasmid transformation event of strain DI-9 was the result of uptake and expression of plasmid DNA by a competent mutant (HfT strain). Additionally, it was found that a strain of Vibrio parahaemolyticus, USFS 3420, could be naturally transformed with plasmid DNA. Natural plasmid transformation by high-transforming mutants may be a means of plasmid acquisition by natural aquatic bacterial populations.

摘要

河口细菌弧菌DI-9菌株已被证明能够自然转化,分别以平均每受体3.5×10⁻⁹和3.4×10⁻⁷转化子的频率,与广泛宿主范围的质粒多聚体和同源染色体DNA进行转化。质粒转化子在非选择性培养基中生长会产生治愈菌株,这些治愈菌株的转化频率比亲本菌株高6至42857倍,具体取决于转化DNA的类型。这些高频转化(HfT)菌株与质粒DNA的转化频率为每受体1.1×10⁻⁸至1.3×10⁻⁴转化子,与同源染色体DNA的转化频率平均为每受体8.3×10⁻⁵转化子。使用携带转座子Tn5的R1162衍生物的多聚体(pQSR50)时观察到最高的转化频率。对其中一个治愈菌株的总DNA制剂进行检测表明,治愈菌株中没有残留可能与转化DNA提供同源性的质粒DNA。所有转化子和治愈菌株都可以通过菌落形态与亲本菌株区分开来。DNA结合研究表明,对数后期的HfT菌株结合[³H]噬菌体λDNA的速度比亲本菌株快2.1倍。这些结果表明,DI-9菌株最初的质粒转化事件是一个感受态突变体(HfT菌株)摄取和表达质粒DNA的结果。此外,还发现副溶血性弧菌USFS 3420菌株能够用质粒DNA进行自然转化。高转化突变体的自然质粒转化可能是天然水生细菌群体获取质粒的一种方式。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f5/184975/5f96307b5924/aem00092-0219-a.jpg

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