Cloning And Abiotic Functional Analysis Of Salt-tolerant Genes In Halophilic Aspergillus Glaucus

Halopgilic microorganisms has many crucial genes which controls the formation of salt-stress tolerant mechanisms, making it possible to be survived under different salt-stress environment. In order to utilize the genetic resources of halophilic microorganisms effectively, it is meaningful of researching the functions of salt-tolerant genes. 26 S protease regulatory subunit 7(S7), located in the 19 S regulatory particle of 26 S proteasome, has conserved domain and involved in the formation of ATPase activity center. 26 S proteasome is involved in many metabolic processes such as mitosis, programmed cell death and DNA damage repair and so on. In the ubiquitin/26 S proteasome pathway of abnormal protein degradation, the ATPase activity center provides energy for the deubiquitin and translocation of proteins. Aederolysins have common features of stability in a wide p H range, applications in medicine. However, there is no reports about abiotic functions of these two genes. In this study, two salt-tolerant genes were isolated from the yeast expression library of halophilic Aspergillus glaucus CCHA and the abiotic function of the genes were analyzed. The results are as following:1. We successfully cloned the whole sequence of S7 from Aspergillus glaucus, and transported it into yeast and Thermomyces lanuginosus. The transgenic yeast and Thermomyces lanuginosus shows no significant ability of resistance of the Na Cl, sorbitol conditions.2. The non-full-length sequence S7(lack 114bp) was cloned from the Aspergillus glaucus and the yeast isolated from the library, which was named as n Ag S7 and NAg S7. The yeast with p YES2-n Ag S7 or p YES2-NAg S7 shows no ability of resisting the Na Cl and sorbitol conditions. However, the yeast isolated from the library shows significant ability of resisting the Na Cl, sorbitol stresses. We speculate that the yeast was mutated during the construction of the library or has other genes with abiotic functions.3. Transgenic yeast and Thermomyces lanuginosus with Ag Aeg S gene show the ability of resisting the Na Cl, sorbitol stresses.