ISOLATION AND SCREENING OF PLASMIDS FROM THE EPILITHON WHICH MOBILIZE RECOMBINANT PLASMID PD10
ISOLATION AND SCREENING OF PLASMIDS FROM THE EPILITHON WHICH MOBILIZE RECOMBINANT PLASMID PD10
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DOI：
10.1128/aem.58.4.1292-1300.1992
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发表时间：
1992-04-01
影响因子：
4.4
通讯作者：
FRY, JC
中科院分区：
文献类型：
--
作者：
HILL, KE;WEIGHTMAN, AJ;FRY, JC
这项研究检验了来自河石的细菌动员重组分解代谢质粒 pD10 的潜力，该质粒编码 3-氯苯甲酸酯降解和卡那霉素抗性。 通过恶臭假单胞菌菌株和来自塔夫河（英国南威尔士）的石生细菌之间的三亲交配，外源分离出 54 个动员质粒。 在 20 摄氏度下，每个接受者的动员频率范围为 1.7 x 10(-8) 至 4.5 x 10(-3)。 分离出的动员质粒的大小从 40 kb 到超过 200 kb 不等，发现 54 个中的 19 个编码耐汞性。 还发现了质粒编码的对四环素和链霉素的抗性，但没有对紫外线或各种重金属的抗性。 通过比较限制性片段化模式对附石细菌的八个质粒进行分析，显示从不同独立交配中分离的质粒之间存在显着差异。 pD10 动员的最佳温度在 15 至 25 摄氏度之间。 发现四种抗汞质粒具有广泛的宿主范围，可将汞抗性转移并轻松地将 pD10 转移到 β 和 γ 紫细菌的代表性物种中。 一般来说，石生细菌的质粒动员 pD10 的频率比配偶转移频率低 2 至 3 个数量级。 因此，通过多种细菌物种之间的动员而引入表皮的重组基因的交换具有很高的潜力。
This study examined the potential of bacteria from river epilithon to mobilize a recombinant catabolic plasmid, pD10, encoding 3-chlorobenzoate degradation and kanamycin resistance. Fifty-four mobilizing plasmids were exogenously isolated by triparental matings between strains of Pseudomonas putida and epilithic bacteria from the River Taff (South Wales, United Kingdom). Frequencies for mobilization ranged from 1.7 x 10(-8) to 4.5 x 10(-3) per recipient at 20-degrees-C. The sizes of the mobilizing plasmids isolated ranged from 40 kb to over 200 kb, and 19 of 54 were found to encode mercury resistance. Plasmid-encoded resistance to tetracycline and streptomycin was also found but not resistance to UV light or various heavy metals. Eight plasmids of epilithic bacteria, analyzed by comparing restriction fragmentation patterns, showed significant differences between those isolated from different independent matings. Optimal temperatures for mobilization of pD10 were between 15 and 25-degrees-C. Four mercury resistance plasmids were found to be broad host range, transferring mercury resistance and mobilizing pD10 readily to representative species of beta- and gamma-purple bacteria. In general, frequencies of pD10 mobilization by plasmids of epilithic bacteria were 2 to 3 orders of magnitude lower than conjugal transfer frequencies. Thus, there is a high potential for exchange of recombinant genes introduced into the epilithon by mobilization between a variety of bacterial species.