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煤炭资源开发战略问题分析
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作者 李通 《内蒙古煤炭经济》 2013年第3期47-47,49,共2页
煤炭是应用较为广泛的能源形式,属于不可再生的战略资源,我国的煤炭能源储备丰富,但随着改革开放以来的大力开发利用,煤炭的储备已经有了大幅度的减少,本文主要探讨了煤炭资源开发过程中的战略手段,提高我国的能源供给质量。
关键词 基地重组 开采精度 生态保护
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Cloning,Expressing,and Hemolysis of tdh,trh and tlh Genes of Vibrio parahaemolyticus 被引量:2
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作者 ZHAO Yonggang TANG Xiaoqian ZHAN Wenbin 《Journal of Ocean University of China》 SCIE CAS 2011年第3期275-279,共5页
Vibrio parahaemolyticus (VP) is one of the pathogenic vibrios endangering net-cage cultured Pseudosciaena crocea,Fennerpenaeus chinensis, and shellfish in coastal areas of China. Several types of hemolysins produced b... Vibrio parahaemolyticus (VP) is one of the pathogenic vibrios endangering net-cage cultured Pseudosciaena crocea,Fennerpenaeus chinensis, and shellfish in coastal areas of China. Several types of hemolysins produced by Vp have been characterized as major virulence factors.They are thermostable direct hemolysin (TDH),TDH-related hemolysin (TRH) and thermolabile hemolysin (TLH). In this study, we cloned tdh, trh, and tlh genes from the genome DNA of VP by polymerase chain reaction (PCR).We ligated the three genes into prokaryotic expression vector pET-28a (+),and transformed the recombinant plasmids into Es-cherichia coli BL21 (DE3). The expression of recombinant proteins was induced by isopropyl-β-D-thiogalacto-pyranoside (IPTG). The recombinant proteins were expressed in a form of inclusion bodies and thus purified with Ni-NTA affinity chromatography. Western blotting results showed that recombinant proteins,TDH, TRH and TLH, could be recognized by rabbit anti-VP serum. The three purified proteins were renatured by gradient dialysis.The renatured proteins exhibited hemolytic activity except for TLH in the presence of phosphatidylcholine. These results not only are helpful for better understanding these genes' functions under a single factor level, but also provide evidence for VP vaccine engineering. 展开更多
关键词 Vibrio parahaemolyticus HEMOLYSIN CLONE expression hemolytic activity
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