Construction of a Recombinant Lac Z Gene in Yeast Cell for Rapid Detection of Tetracycline Antibiotics_Journal of Biomedical Engineering

Authors：

XUXiuju ¹ , TIANYongjie ¹ , WANGChao ¹ , ZHOUTianqi ¹ , ZHOULuojing ² ,  LIXiangming ¹

1. Preventive Medicine Department, Yangzhou Medical College, Yangzhou University, Yangzhou 225001, China;
2. Subei People's Hospital, Yangzhou 225001, China;

Corresponding author：

LIXiangming, Email: 847264344@qq.com

Keywords：

yeast cells; tetracycline; tetracycline repressor protein; Lac Z gene

DOI：

10.7507/1001-5515.20160081

Video：

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Two vectors were used to construct the recombinant gene yeast cell that can be used to bioassay of the pollution of tetracycline antibiotics in the environment. In the expression vector, the GPD (glyceraldehyde-3-phosphate dehydrogenase) promoter was used to drive the gene expression of tetracycline repressor protein (TR) fused with V5 antigen epitope gene, while in the reporter vector, the tetracycline response element (TRE) was used to regulate Lac Z report gene expression. The specificity and the sensitivity of the recombinant gene yeast cell were evaluated respectively by different concentrations of tetracycline antibiotics and non-tetracycline antibiotics. The results showed that there were significant dose effect relationships between the tetracycline antibiotics and the yeast cells, while non-tetracycline antibiotics showed no dose effect relationships with this biosensor. It is illustrated that the recombinant yeast cells can be used to monitor the tetracycline antibiotic pollution on the environment.

Citation： XUXiuju, TIANYongjie, WANGChao, ZHOUTianqi, ZHOULuojing, LIXiangming. Construction of a Recombinant Lac Z Gene in Yeast Cell for Rapid Detection of Tetracycline Antibiotics. Journal of Biomedical Engineering, 2016, 33(3): 481-487. doi: 10.7507/1001-5515.20160081 Copy

1.	刘雨霞,鲍艳宇.土壤中四环素类抗生素污染研究进展[J].环境污染与防治,2011,33(8) :81-86, 91.
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8.	吕海涛,陈峰,姜悦.高效液相色谱法测定四环素类抗生药物中土霉素、强力霉素、四环素和金霉素[J].理化检验(化学分册),2007,43(1) :18-20.
9.	刘虹,张国平,刘丛强.固相萃取-色谱测定水、沉积物及土壤中氯霉素和3种四环素类抗生素[J].分析化学,2007,35(3) :315-319.
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1. 刘雨霞,鲍艳宇.土壤中四环素类抗生素污染研究进展[J].环境污染与防治,2011,33(8) :81-86, 91.
2. 张肃川,梁定伟,何小平.四川省细菌耐药监测网2011年川南地区细菌耐药性监测数据分析[J].实用医院临床杂志,2012,9(6) :115-120.
3. RATASUK N, BOONSANER M, HAWKER D W. Effect of temperature, pH and illumination on abiotic degradation of oxytetracycline in sterilized swine manure[J]. J Environ Sci Health A Tox Hazard Subst Environ Eng, 2012, 47(11) :1687-1694.
4. WANG Qiquan, YATES S R. Laboratory study of oxytetracycline degradation kinetics in animal manure and soil[J]. Food Chem, 2008, 56(5) :1683-1688.
5. CHEE-SANFORD J C, MACKIE R I, KOIKE S A, et al. Fate and transport of antibiotic residues and antibiotic resistance genes following land application of manure waste[J]. J Environ Qual, 2009, 38(3) :1086-1108.
6. HILLEN W, BERENS C. Mechanisms underlying expression of Tn10 encoded tetracycline resistance[J]. Annu Rev Microbiol, 1994, 48:345-369.
7. MEIER I, WRAY L V, HILLEN W. Differential regulation of the Tn10-encoded tetracycline resistance genes tetA and tetR by the tandem tet operators O1 and O2[J]. EMBO J, 1988, 7(2) :567-572.
8. 吕海涛,陈峰,姜悦.高效液相色谱法测定四环素类抗生药物中土霉素、强力霉素、四环素和金霉素[J].理化检验(化学分册),2007,43(1) :18-20.
9. 刘虹,张国平,刘丛强.固相萃取-色谱测定水、沉积物及土壤中氯霉素和3种四环素类抗生素[J].分析化学,2007,35(3) :315-319.
10. 张慧敏,章明奎,顾国平.浙北地区畜禽粪便和农田土壤中四环素类抗生素残留[J].生态与农村环境学报,2008,24(3) :69-73.
11. KORPELA M T, KURITTU J S, KARVINEN J T, et al. A recombinant Escherichia coli sensor strain for the detection of tetracyclines[J]. Anal Chem, 1998, 70(21) :4457-4462.

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Construction of a Recombinant Lac Z Gene in Yeast Cell for Rapid Detection of Tetracycline Antibiotics

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