Application of Green Fluorescent Protein as Gene Marker for Microbial Enhanced Oil Recovery

doi:10.3696/j.issn.1006-396X.2009.04.009

Journal of Petrochemical Universities ›› 2009, Vol. 22 ›› Issue (4): 34-37.DOI: 10.3696/j.issn.1006-396X.2009.04.009

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Application of Green Fluorescent Protein as Gene Marker for Microbial Enhanced Oil Recovery

Faculty of Chemical Science and Engineering, China University of Petroleum, Beijing 102249, P.R.China

Received:2008-03-31 Revised:2009-04-15 Published:2009-12-25 Online:2009-12-25

绿色荧光蛋白标记在MEOR中的应用研究

张忠智,孙珊珊,王欲晓,罗一菁

中国石油大学(北京)化工学院

作者简介:张忠智(1955 -), 男, 山东嘉祥县, 教授, 博士
基金资助:
中国石油天然气股份有限公司科技风险创新研究项目(2005Petrichina -develo p.02);国家973 项目(2005cb221308)

Abstract

Abstract:

In order to obtain more analyses and estimate correctly and accurately to improve Microbial Enhanced Oil Recovery (MEOR), goal strains for MEOR would be marked and tracked to indicate that how and where they moved and reproduced in oil reservoir and show their amount and rate in oil-field microorganism. Recombinant plasmid of pET-30a(+)-EGFP which has the gene of green fluorescent protein (GFP) was constructed and transferred into JD successfully . Recombinant plasmid can be stably maintained in the culture media including 50 μg/mL kanamycin (Kan), which indicated recombinant plasmid stability under choice pressure, while without kanamycin it can be lost easily. JD(gfp) can normally reproduce and metabolize together with wild strain for MEOR isolated from oil reservoir while no transfer of recombinant plasmid.

Key words: Microbial enhanced oil recovery(MEOR) , Green fluorescent protein(GFP) , Gene marker , Recombinant plasmid , Strain JD

摘要： 为了充分认识目的优势菌种经油藏运移后在产出水中分布的规律与数量,准确分析和客观评价MEOR现场试验效果,利用绿色荧光蛋白(GFP)基因标记目的菌,通过绿色荧光蛋白跟踪监测目的菌的动态信息。构建含有GFP基因的质粒pET-30a(+)-EGFP,并将其成功导入JD中。对构建的JD(gfp)工程菌进行培养发现,该工程菌在含有Kan质量浓度50μg/mL的培养条件下,重组质粒在JD(gfp)工程菌体内传代稳定,绿色荧光的表达稳定。但是在没有选择压力的培养条件下,重组质粒在JD工程菌胞体内有丢失现象。与野生菌混合培养,JD(gfp)工程菌具有很好的配伍性和竞争性,并且不发生重组质粒的水平转移,因此在环境中释放是安全的。

关键词: 微生物采油 , 绿色荧光蛋白(GFP) , 基因标记 , 重组质粒 , JD菌

ZHANG Zhong-zhi, SUN Shan-shan, WANG Yu-xiao，et al. Application of Green Fluorescent Protein as Gene Marker for Microbial Enhanced Oil Recovery[J]. Journal of Petrochemical Universities, 2009, 22(4): 34-37.

张忠智,孙珊珊,王欲晓,罗一菁. 绿色荧光蛋白标记在MEOR中的应用研究[J]. 石油化工高等学校学报, 2009, 22(4): 34-37.

Application of Green Fluorescent Protein as Gene Marker for Microbial Enhanced Oil Recovery

绿色荧光蛋白标记在MEOR中的应用研究

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