PAN Zhuo

doi:10.3967/bes2016.030

Optimization of Pulsed-field Gel Electrophoresis Procedure for Bacillus cereus

文章导航 > Biomedical and Environmental Sciences > 2016 > 29(3): 233-238

PAN Zhuo. Optimization of Pulsed-field Gel Electrophoresis Procedure for Bacillus cereus[J]. Biomedical and Environmental Sciences, 2016, 29(3): 233-238. doi: 10.3967/bes2016.030

Citation:

PAN Zhuo. Optimization of Pulsed-field Gel Electrophoresis Procedure for Bacillus cereus[J]. Biomedical and Environmental Sciences, 2016, 29(3): 233-238. doi: 10.3967/bes2016.030

doi: 10.3967/bes2016.030

基金项目: This study was supported by the National Special Fund for the Development of Major Research Equipment and Instruments (2012YQ09019706)from the Ministry of Science and Technology, and by the Priority Project on Infectious Disease Control and Prevention (2012ZX10004215 and 2008ZX10004-008) from the National Health and Family Planning Commission of the Peoples Republic of China

Optimization of Pulsed-field Gel Electrophoresis Procedure for Bacillus cereus

PAN Zhuo

Microbiology Laboratory, Shijiazhuang Center for Disease Control and Prevention, Shijiazhuang 050016, Hebei, China

Funds: This study was supported by the National Special Fund for the Development of Major Research Equipment and Instruments (2012YQ09019706)from the Ministry of Science and Technology, and by the Priority Project on Infectious Disease Control and Prevention (2012ZX10004215 and 2008ZX10004-008) from the National Health and Family Planning Commission of the Peoples Republic of China

摘要: In order to develop a rapid and reliable method forB. cereusgenotyping, factors influencing PFGE results, including preparation of bacterial cells embedded in agarose, lysis of embedded cells, enzymatic digestion of intact genomic DNA, and electrophoresis parameters allowing for reproducible and meaningful DNA fragment separation, were controlled. Optimal cellular growth (Luria-Bertani agar plates for 12-18 h) and lysis conditions (4 h incubation with 500 μg/mL lysozyme) produced sharp bands on the gel. Restriction enzymeNotI was chosen as the most suitable. Twenty-two isolates were analyzed by NotI digestion, using three electrophoretic parameters (EPs). The EP-a was optimal for distinguishing between isolates. The optimized protocol could be completed within 40 h which is a significant improvement over the previous methods.
- /
- /
- Bacillus cereus /
- DNA fragment /
- genomic DNA

[1]	YAN Zhao Fan, GU Zhi Guang, FAN Ya Hui, LI Xin Ling, NIU Ze Ming, DUAN Xiao Ran, Mallah Ali Manthar, ZHANG Qiao, YANG Yong Li, YAO Wu, WANG Wei. Benchmark Dose Assessment for Coke Oven Emissions-Induced Mitochondrial DNA Copy Number Damage Effects . Biomedical and Environmental Sciences, 2023, 36(6): 490-500. doi: 10.3967/bes2023.060
[2]	PAN Dong, DU Ya Rong, LI Rong, SHEN Ai Hua, LIU Xiao Dong, LI Chuan Yuan, HU Bu Rong. SET8 Inhibition Potentiates Radiotherapy by Suppressing DNA Damage Repair in Carcinomas . Biomedical and Environmental Sciences, 2022, 35(3): 194-205. doi: 10.3967/bes2022.028
[3]	ZHANG Wen Ping, ZHANG Yi Fan, ZHANG Ying Ying, HAN Zhi Chao, GAO Yuan Yuan, GUO Jian Yong, SHI Xiu Jing, HU Xiao Qin, MU Li Na, ZHOU Yun, LEI Li Jian. Peripheral Blood Mitochondrial DNA Copy Number and Hypertension Combined with Albuminuria in Chinese Coal Miners . Biomedical and Environmental Sciences, 2021, 34(7): 567-571. doi: 10.3967/bes2021.078
[4]	YUKSEL Selin, DINÇER Yildiz. Relationship between Exposure to Low Dose of x-ray and DNA Hypomethylation in Solid Tumors and Hematological Malignancies . Biomedical and Environmental Sciences, 2020, 33(7): 528-537. doi: 10.3967/bes2020.069
[5]	WU Jian Hui, CHEN Jiao, WANG Yong, XIA Bin, WANG Rong, ZHAO Yan, WANG Qin Xia, SONG Qi, YAO Shun Heng, ZHANG Yun Hui, SUN Zu Yue. Effect of Mono-2-ethyhexyl Phthalate on DNA Methylation in Human Prostate Cancer LNCaP Cells . Biomedical and Environmental Sciences, 2017, 30(9): 641-648. doi: 10.3967/bes2017.085
[6]	LI RuiJing, GAO Hui, NA GuangShui, LU ZiHao, YAO Yao, YANG Fan. Hexabromocyclododecane-induced Genotoxicity in Cultured Human Breast Cells through DNA Damage . Biomedical and Environmental Sciences, 2017, 30(4): 296-300. doi: 10.3967/bes2017.039
[7]	MAO Li Li, XIAO Xin Hua, ZHANG Qian, ZHENG Jia, LI Wen Hui, YU Miao, ZHANG Hua Bing, PING Fan, XU Jian Ping, WANG Xiao Jing. DNA Methylation and Birth Weight:a Genome-wide Analysis . Biomedical and Environmental Sciences, 2017, 30(9): 667-670. doi: 10.3967/bes2017.088
[8]	LING Xiao Xuan, LIU Jia Xian, YUN Lin, DU Yu Jun, CHEN Shao Qian, CHEN Jia Long, TANG Huan Wen, LIU Lin Hua. Poly(ADP-ribosyl)ation of Apoptosis Antagonizing Transcription Factor Involved in Hydroquinone-Induced DNA Damage Response . Biomedical and Environmental Sciences, 2016, 29(1): 80-84. doi: 10.3967/bes2016.008
[9]	YU Gang, WANG Xun Cheng, TIAN Wang Hong, SHI Ji Chun, WANG Bin, YE Qiang, DONG Si Guo, ZENG Ming, WANG Jun Zhi. Genomic Diversity and Evolution of Bacillus subtilis . Biomedical and Environmental Sciences, 2015, 28(8): 620-625. doi: 10.3967/bes2015.087
[10]	XIE Lei, WANG YingWei, GUAN ShanYue, XIE LiJing, LONG Xin, SUN ChengYe. Prospects and Problems for Identification of Poisonous Plants in China using DNA Barcodes . Biomedical and Environmental Sciences, 2014, 27(10): 794-806. doi: 10.3967/bes2014.115
[11]	LIU Qing, GU Jian Hong, YUAN Yan, LIU Xue Zhong, WANG Ya Jun, WANG Han Dong, LIU Zong Ping. Effect of Cadmium on Rat Leydig Cell Testosterone Production and DNA Integrity in vitro . Biomedical and Environmental Sciences, 2013, 26(9): 769-773. doi: 10.3967/0895-3988.2013.09.009
[12]	ZHAO Xiao Tao, FENG Jiang Bin, LI Yu Wen, LUO Qun, YANG Xin Chun, LU Xue, CHEN De Qing, LIU Qing Jie. Identification of Two Novel Mitochondrial DNA Deletions Induced by Ionizing Radiation . Biomedical and Environmental Sciences, 2012, 25(5): 533-541. doi: 10.3967/0895-3988.2012.05.006
[13]	YUAN-HAI YOU, PENG WANG, YAN-HUA WANG, HAI-BIN WANG, DONG-ZHENG YU, RONG HAI, JIAN-ZHONG ZHANG. Assessment of Comparative Genomic Hybridization Experiment by an in situ Synthesized CombiMatrix Microarray with Yersinia pestis Vaccine Strain EV76 DNA . Biomedical and Environmental Sciences, 2010, 23(5): 384-390.
[14]	JUN-WEN LI, JIN-LAI ZHENG, XIN-WEI WANG, MIN JIN, FU-HUAN CHAO. Directed Molecular Evolution of Nitrite Oxido-reductase by DNA-shuffling . Biomedical and Environmental Sciences, 2007, 20(2): 113-118.
[15]	HONG-ZHI PAN, DONG CHANG, LEI-GUANG FENG, FENG-JUAN XU, HONG-YU KUANG, MING-JUN LU. Oxidative Damage to DNA and Its Relationship With Diabetic Complications . Biomedical and Environmental Sciences, 2007, 20(2): 160-163.
[16]	XIAO-HONG ZHAO, GUANG JIA, YONG-QUAN LIU, SHAO-WEI LIU, LEI YAN, YU JIN, NIAN LIU. Association Between Polymorphisms of DNA Repair Gene XRCC1 and DNA Damage in Asbestos-Exposed Workers . Biomedical and Environmental Sciences, 2006, 19(3): 232-238.
[17]	K.KRISHNAMURTHI, S. SARAVANA DEVI, T. CHAKRABARTI. DNA Damage Caused By Pesticide-contaminated Soil . Biomedical and Environmental Sciences, 2006, 19(6): 427-431.
[18]	QUN-FENG YAO, XIN-JIANG KANG, QIAO-LING HAO, Yi-Kai Zhou. A Quantitative DNA Methylation Assay Using Mismatch Hybridization and Chemiluminescence . Biomedical and Environmental Sciences, 2005, 18(1): 48-52.
[19]	WEI ZHENG, JI-LIANG HE, LI-FEN JIN, JIAN-LIN LOU, BAO-HONG WANG. Assessment of Human DNA Repair (NER) Capacity With DNA Repair Rate (DRR) by Comet Assay . Biomedical and Environmental Sciences, 2005, 18(2): 117-123.
[20]	AI-GUO WANG, TAO Xia, QI-LONG CHU, Ming Zhang, FANG LIU, XUE-MIN CHEN, KE-DI YANG. Effects of Fluoride on Lipid Peroxidation, DNA Damage and Apoptosis in Human Embryo Hepatocytes . Biomedical and Environmental Sciences, 2004, 17(2): 217-222.

点击查看大图

计量

文章访问数: 1078
HTML全文浏览量: 311
PDF下载量: 71
被引次数: 0

Optimization of Pulsed-field Gel Electrophoresis Procedure for Bacillus cereus

doi: 10.3967/bes2016.030

PAN Zhuo

刊出日期: 2016-03-20

关键词:

Bacillus cereus /

DNA fragment /

genomic DNA

摘要: In order to develop a rapid and reliable method forB. cereusgenotyping, factors influencing PFGE results, including preparation of bacterial cells embedded in agarose, lysis of embedded cells, enzymatic digestion of intact genomic DNA, and electrophoresis parameters allowing for reproducible and meaningful DNA fragment separation, were controlled. Optimal cellular growth (Luria-Bertani agar plates for 12-18 h) and lysis conditions (4 h incubation with 500 μg/mL lysozyme) produced sharp bands on the gel. Restriction enzymeNotI was chosen as the most suitable. Twenty-two isolates were analyzed by NotI digestion, using three electrophoretic parameters (EPs). The EP-a was optimal for distinguishing between isolates. The optimized protocol could be completed within 40 h which is a significant improvement over the previous methods.

English Abstract

PAN Zhuo. Optimization of Pulsed-field Gel Electrophoresis Procedure for Bacillus cereus[J]. Biomedical and Environmental Sciences, 2016, 29(3): 233-238. doi: 10.3967/bes2016.030

Citation:

PAN Zhuo. Optimization of Pulsed-field Gel Electrophoresis Procedure for Bacillus cereus[J]. Biomedical and Environmental Sciences, 2016, 29(3): 233-238. doi: 10.3967/bes2016.030