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目的为分析我国弯曲菌遗传特征,本研究根据已发表多株弯曲菌的全基因组测序特征及比对结果自行设计基因芯片,利用芯片对我国不同宿主来源菌株进行遗传特异性分析。方法根据前期基因组水平比对分析的结果,利用Combimatrix tiling Custom ArrayTM90K芯片,设计DNA芯片。芯片包含已测序菌株ICDCCJ07001、269.97、NCTC11168、81-176、81-116和RM1221共3384个CDS的探针序列,以及空肠弯曲菌耐药及致病性相关2个质粒共80个CDS的探针序列,与脂寡糖的合成相关基因簇16种共219个CDS的探针序列、荚膜多糖合成相关基因簇7种共160个CDS的所有序列。对我国不同宿主来源27株分离菌株提取DNA,利用芯片进行杂交,获得杂交信息并分析不同菌株CDS分布特征分析及聚类特点。结果中国菌株的主要变异区域主要存在于与脂寡糖、荚膜多糖合成相关的基因簇、鞭毛修饰相关的基因簇、DNA限制/修饰相关的基因簇以及空肠弯曲菌Mu样噬菌体基因簇。基因组水平不同来源菌株CDS分布的聚类结果没有发现显著的宿主归因特点,但GBS相关菌株脂寡糖合成相关基因组成具有共性特征。结论通过验证以及与过去研究的比较,本次研究中的基因芯片技术结果准确可信,本研究所用基因芯片在分析空肠弯曲菌基因多态性方面具有很好的优势,可用于弯曲菌遗传特征和重要毒力因子的分析和检测。
Objective To analyze the genetic characteristics of Campylobacter in our country, based on the genome-wide sequencing features and the comparison results of many strains of Campylobacter strains, we designed the gene chip and used the chip to analyze the genetic diversity of different host strains in our country. Methods Based on the results of previous analysis of genomic levels, DNA chips were designed using Combimatrix tiling Custom ArrayTM 90K chip. The chip contains 3384 CDS probe sequences of the sequenced strains ICDCCJ07001, 269.97, NCTC11168, 81-176, 81-116 and RM1221, as well as 80 CDS probes of two plasmids resistant to Campylobacter jejuni and pathogenicity Sequence, a total of 219 CDS probe sequences of 16 gene clusters related to the synthesis of lipooligosaccharides, and all seven sequences of 160 CDS sequences of 7 capsular polysaccharide synthesis related genes. DNA was extracted from 27 strains of isolates from different host countries in our country, and the hybridization was performed by using the chip. The hybridization information was obtained and the CDS distribution characteristics and clustering characteristics of different strains were analyzed. Results The main variation regions of Chinese strains were found in the gene cluster related to lipooligosaccharides and capsular polysaccharide synthesis, flagella modification related gene cluster, DNA restriction / modification related gene cluster and Campylobacter jejuni Mu-like phage gene cluster. No significant host-attribution was found in the clustering results of CDS distributions of different strains of genomic level, but the gene composition related to lipo-oligosaccharide synthesis of GBS-related strains had common features. Conclusion Through the verification and comparison with the past research, the gene chip technology in this study is accurate and credible. The gene chip used in this study has a good advantage in analyzing the genetic polymorphism of Campylobacter jejuni and can be used for the genetic characteristics of Campylobacter And analysis and detection of important virulence factors.