Physiological Function of Membrane Protein RHOGL009301 Involved in Transport of Benzoate in Rhodococcus sp. R04
Acta Microbiologica Sinica,Vol 57,No. 04
[Objective] The physiological functions of membrane protein RHOGL009301 in Rhodococcus sp. R04 and the metabolic properties of the mutant strain were studied to determine the relationship between the physiological function of the membrane protein and the transport of benzoate. [Methods] The RHOGL009301 gene and the green fluorescent protein gene were fused for expressing in Rhodococcus erythropolis, and the location of RHOGL009301 was observed by Delta Vision. The RHOGL009301 gene was knocked out by homologous recombination, and the growth of wild strain and deficient strain in different carbon sources were compared. The intracellular and extracellular metabolites of the wild strain and the deficient strain when grew on biphenyl and benzoate were measured by HPLC, and the changes of metabolite concentration in different growth conditions were analyzed. [Results] A fusion gene that contained RHOGL009301 gene and the green fluorescent protein gene was co-expressed in Rhodococcus erythropolis and localized on the cell membrane. The deficient strain R04ΔMP of RHOGL009301 gene was obtained. The biomass of the deficient strain was significantly reduced in biphenyl and benzoate culture, and its growth rate was decreased. HPLC analysis showed that the deletion of RHOGL009301 gene inhibited the transport of benzoate. [Conclusion] RHOGL009301 membrane protein is one of the proteins involved in metabolism and transport of benzoate. Based on sequence homology analysis, we can conclude that the membrane protein is a novel benzoate transport protein.
Gene cloning, polyclonal antibody preparation and expression localization of the endophilins from Nilaparvata lugens (Hemiptera: Delphacidae)
Acta Entomologica Sinica,Vol 60,No. 03
[Aim] To explore the function of endophilin in the growth and reproduction of the brown planthopper, Nilaparvata lugens. [Methods] The endophilin genes of N. lugens were cloned and analyzed by bioinformatics. Then, recombinant prokaryotic expression vectors were constructed and transferred into Escherichia coli Rosetta to induce the expression of fusion proteins. After purification by Ni affinity chromatography, the fusion proteins were used to immunize the New Zealand rabbits to obtain polyclonal antibodies against endophilins. The obtained antibodies were used to detect the expression and localization of endophilins in N. lugens ovary. [Results] Two endophilin genes, endophilin A ( Endo A) and endophilin B ( Endo B) , were cloned from N. lugens, with the GenBank accession numbers of KY126096 and KY126095, respectively. Endo A and Endo B encode 387 and 352 amino acids, respectively. Both encoded proteins contain the typical BAR domain and SH3 domain, but show different structure. The titer of the obtained antibodies was estimated as high as 1:1, 000, 000 dilution ratio through ELISA, and the antibodies had good specificity as shown by Western blot. Using immunofluorescence, the antibodies were used to detect the expression of Endo A and Endo B in N. lugens ovary. The results indicated that Endo A and Endo B were universally expressed in N. lugens ovary. In ovary, Endo A and Endo B were widely distributed in the extracellular space, cytomembrane and cytoplasm of follicle cells of N. lugens ovary, and this distribution pattern was similar to that of lipids. Meanwhile, Endo A and Endo B were colocalized with yeast-like symbionts invading into N. lugens ovary. [Conclusion] The nucleotide sequences and the biological characteristics of Endo A and Endo B were clarified. With the obtained polyclonal antibodies of Endo A and Endo B, the expression of Endo A and Endo B in N. lugens ovary was profiled, and the results suggest that Endo A and Endo B might be associated with the development and maturity of N. lugens ovary as well as the invasion of the yeast-like endosymbionts to N. lugens ovary. These results lay the foundation for further studies on the biological function of Endo A and Endo B in N. lugens.
Northeast Asia Forum,Vol 26,No. 03
Security and cooperation should be a serious problem for East Asia which becomes more intense and more complex as a result of the solidification of the dualistic structure in this region since the end of the Cold War. Although this situation leaves many barriers for the Belt and Road Initiative, at the same time, it may provide the countries involved with opportunities and resources to reconstruct the security and cooperation regime. Moreover, this regime should be a good match for the Belt and Road Initiative by reconstructing strategic objectives, frameworks, functions, organizations and principles. Simultaneously, it should be the most effective way to remove the source of major obstacles to security and corporation in this region.
Social memory’s restoration and reconstruction in the reconstruction of disaster-stricken areas: taking Shanghai social work empowerment service in earthquake-stricken Ludian as an example
Sociological Studies,Vol 31,No. 02
During the reconstruction process of disaster-stricken areas, the essence of empowering social memory is the empowerment of innate power and endogenous capacity. With the professional social work services by Shanghai social work team in earthquake-stricken Ludian as an example, the empowerment method in professional social work is explored and its possibility of interventions in disasters is evaluated. This paper analyzes service projects operated at the individual psychological level, interpersonal level, and the level of community organizations and their impact on the disaster restoration and reconstruction of social memories. It further investigates the path and prospect of involving the professional social work empowerment method in affected areas.
Sociological Studies,Vol 31,No. 04
In the mid-1930s, Wu Wenzao combined human ecology and ethnography to create an influential methodology of the “community study.” In this article, the author reconsiders Wu’s synthesis. As Wu emphasizes, rural communities are “life worlds” in the broad sense of the term. They are sites where the relationships between persons, persons and things, and persons and divinities (sometimes overlapping with persons and things) co-exist. These sites are parts of the larger regional and civilizations whole, but they themselves are also wholes, being places where old and new traditions come to be re-engaged in the local life. This is the very essence of the “little tradition.” Wu rightly defined the community as a world of material (ecological), social, and “spiritual” existence. However, being a social-centric sociologist, he did not allow an adequate space for the understanding of the material and “spiritual” aspects of the “life world” of the community. In this methodological critique, the author reflects on the existing theories of the “little” and “great” traditions.