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Abstract：
The objective of this study was to identify genes associated with the biodegradation of phenol by Acinetobacter sp. strain DF4 through the use of differential display (DD) methodology. The bacteria were grown in YEPG medium, and total RNA was extracted and analyzed using labeled primers to detect gene expression differences. Three distinctively expressed cDNA bands (ph1, ph2, and ph3) were identified, cloned, and sequenced. DNA analysis involved searching for open reading frames (ORFs), verifying results with the NCBI database, predicting promoter regions, and constructing phylogenetic trees using bioinformatics tools. The ph1 gene displayed a 97% identity with the AraC transcriptional regulator, suggesting its potential role in regulating the ortho-catabolic pathway of phenol. The ph2 gene showed a 98% identity with aspartate semialdehyde dehydrogenase, which is involved in phenol degradation. The ph3 gene had a 93% identity with acetyltransferase. Essential transcription factors, such as TATA, GTGTGT, CACA, and CTTTT, were detected, and the three genes promoter regions were predicted. This study successfully identified functional genes involved in the metabolism of cyclic chemicals, particularly phenol, using the DD technique. These findings provide insights into the biodegradation pathways of phenol by Acinetobacter sp. Strain DF4 and may contribute to the development of more efficient bioremediation strategies for phenol-contaminated environments.
摘要：
这项研究的目的是鉴定与不动杆菌属生物降解苯酚相关的基因。通过使用差分显示(DD)方法对DF4进行应变。细菌在YEPG培养基中生长，和总RNA提取和分析使用标记的引物来检测基因表达差异。鉴定出三个独特表达的cDNA条带（ph1，ph2和ph3），克隆,并测序。DNA分析涉及搜索开放阅读框(ORF)，用NCBI数据库验证结果，预测启动子区域，利用生物信息学工具构建系统发育树。ph1基因与AraC转录调节因子具有97％的同一性，提示其在调节苯酚的邻位分解代谢途径中的潜在作用。ph2基因与天冬氨酸半醛脱氢酶有98%的同一性,参与苯酚降解。ph3基因与乙酰转移酶具有93%的同一性。必需转录因子，比如TATA,GTGTGT,CACA,和CTTTT,被检测到，并对这三个基因的启动子区域进行了预测。这项研究成功地确定了参与环状化学物质代谢的功能基因，特别是苯酚，使用DD技术。这些发现为不动杆菌对苯酚的生物降解途径提供了见解。菌株DF4和可能有助于为苯酚污染环境开发更有效的生物修复策略。
