新记录:铜绿假单胞菌Rhamnolips(rhlA)基因的分子表达

IF 0.7 Q4 MICROBIOLOGY Journal of Pure and Applied Microbiology Pub Date : 2023-09-01 DOI:10.22207/jpam.17.3.25
F. N. Al-Kanany, Najim Mohsen
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引用次数: 0

摘要

土壤样本采集于伊拉克巴士拉库尔纳西部受石油污染的地点。假单胞菌最初在矿物盐和假单胞菌琼脂培养基上分离,并通过形态和生化表征进行鉴定。然后,根据引物设计条件设计属于铜绿假单胞菌的rhlA基因的特异性引物,并进行PCR扩增得到大小为888 bp的rhlA基因片段;此外,纯化一次PCR产物并送去测序。在凝胶上确定了约888bp的条带,对扩增的rhlA基因测序结果进行了修正,仅366 bp可以使用(BLAST)软件进行分析,最终结果被鉴定为与铜绿假单胞菌相关的染色体rhlA基因部分序列,鉴定率为99.45%。查询基因与NCBI上另一个部分rhlA记录的不完全匹配是由两个碱基对序列(分别为序列348中的T和序列353中的C)的差异引起的,尽管差异很小,但这导致了所产生的氨基酸的差异;以便在将序列存入GenBank时分配一个新的记录号ON637169。通过建立系统发育树,证实了新记录的部分rhlA基因与其他60条相同数量的rhlA基因序列之间的关系。
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New Record: Molecular Depiction of Rhamnolipids (rhlA) Gene in Locally Isolated Strains of Pseudomonas aeruginosa
Soil samples were collected from oil-contaminated sites which were located in west Qurna, Basrah, Iraq. Pseudomonas species were initially isolated on mineral salts and Pseudomonas agar media and identified using morphological and biochemical characterizations. Then, specific primers for the rhlA gene belonging to Pseudomonas aeruginosa were designed based on the primer design conditions, and PCR was performed to amplify the 888 bp size fragment of the rhlA gene; additionally, the primary PCR products were purified and sent for sequencing. The band of about 888bp was determined on the gel, the amplified rhlA gene sequencing findings were revised, only 366 bp were ready to analyze using the (BLAST) software, and the final result was identified as a partial sequence of chromosomal rhlA gene related to Pseudomonas aeruginosa with percent identity of 99.45%. The query gene’s incomplete matching with another partial rhlA record on NCBI was caused by variations in two base pair sequences (T in sequence 348 and C in sequence 353, respectively), and despite the small difference, this results in variation in the amino acids produced; so that a new record number, ON637169, was assigned when the sequence was deposited in GenBank. The relation among the new record of partial rhlA gene with the same number of the other rhlA gene sequences (60 records) was demonstrated by creating a phylogenetic tree.
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来源期刊
Journal of Pure and Applied Microbiology
Journal of Pure and Applied Microbiology BIOTECHNOLOGY & APPLIED MICROBIOLOGY-MICROBIOLOGY
CiteScore
2.00
自引率
0.00%
发文量
266
审稿时长
11 months
期刊介绍: Journal of Pure and Applied Microbiology (JPAM) is a peer-reviewed, open access international journal of microbiology aims to advance and disseminate research among scientists, academics, clinicians and microbiologists around the world. JPAM publishes high-quality research in all aspects of microbiology in both online and print form on quarterly basis.
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