Cloning of the major protein of the Caulobacter crescentus periodic surface layer: detection and characterization of the cloned peptide by protein expression assays
Cloning of the major protein of the Caulobacter crescentus periodic surface layer: detection and characterization of the cloned peptide by protein expression assays
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新月柄杆菌周期性表面层主要蛋白质的克隆：通过蛋白质表达测定对克隆肽进行检测和表征
DOI：
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发表时间：
1984
影响因子：
3.2
通讯作者：
Nina Agabiant
中科院分区：
文献类型：
--
作者：
J. Smitt;Nina Agabiant
在细菌Caulobacter crescentus CB 15的表面发现了精确有序的晶体阵列。利用免疫学分析，我们从C. crescentus CB15 DNA.一个单一的4.4-丝氨酸蛋白酶HindIII片段编码的多肽，其抗原决定簇，分子量和特有的溶解特性是与那些真正的主要多肽（130 K）的表面阵列相同。130 K蛋白作为从λ启动子开始的基因转录的结果而作为离散产物产生;几个实验表明，该基因的柄杆菌属启动子不能被大肠杆菌转录机器有效地识别。基因组Southern分析揭示了每个基因组中130 K蛋白基因的单拷贝。将130 K蛋白基因与两株关系密切的实验室分离株的DNA进行杂交。crescentus已经失去了产生表面阵列的能力。这些菌株之一，CB 2，具有130 K基因的同源拷贝，而从其他菌株，CB 13 B1 a，DNA显示出较低程度的杂交到130 K基因探针;杂交的基因组片段是不同的大小相比，在CB 13的CB 15。这些研究结果进行了讨论有关的表面阵列的功能和它在细胞形态发生中的作用，在这个茎形成细菌的研究。
A precisely ordered crystalline array is found on the surface of the bacterium Caulobacter crescentus CB15. Using an immunological assay, we identified recombinant bacteriophage clones expressing the predominant protein of this structure from a lambda 1059 library of C. crescentus CB15 DNA. A single 4.4-kilobase HindIII fragment encoded a polypeptide whose antigenic determinants, molecular weight, and peculiar solubilization properties were identical with those of the authentic predominant polypeptide (130K) of the surface array. The 130K protein was produced as a discrete product as a result of gene transcription initiated from a lambda promoter; several experiments suggested that the Caulobacter promoter for this gene is not efficiently recognized by the Escherichia coli transcription machinery. Genomic Southern analysis revealed a single copy of the 130K protein gene per genome. The 130K protein gene was hybridized with DNA of two closely related laboratory strains of C. crescentus which have lost their ability to produce a surface array. One of these strains, CB2, possesses an homologous copy of the 130K gene, whereas DNA from the other strain, CB13B1a, showed a lesser degree of hybridization to the 130K gene probe; genomic fragments which did hybridize were of different sizes in CB13 as compared with those of CB15. These findings are discussed in relation to studies of the surface array function and its role in cellular morphogenesis in this stalk-forming bacterium.
新月柄杆菌钩蛋白基因克隆的分离和表达。
DOI：
10.1073/pnas.79.16.4863
发表时间：
1982
影响因子：
11.1
作者：
Ohta,N;Chen,LS;Newton,A
通讯作者：
Newton,A