http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
Establishment and Application of Target Gene Disruption System in Saccharomyces boulardii
Longjiang Wang,Hui Sun,Jie Zhang,Qing Liu,Tiantian Wang,Peipei Chen,Hongmei Li,Yihong Xiao,Fangkun Wang,Xiaomin Zhao 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.1
Saccharomyces boulardii is the best knownprobiotic yeast, widely used as a therapeutic agent for thetreatment or prevention of diarrhea and intestine disorders. In the present work, we established a target gene disruptionsystem for S. boulardii based on the Cre-loxP system usedfor S. cerevisiae and other fungi by screening out selectionmarkers, working out the transformation method, andconstructing essential plasmids for S. boulardii. Theestablished system was successfully applied to the URA3gene disruption and created an ura3 null mutant strain ofS. boulardii. The system can be used for PCR mediatedgene disruption, cloning mediated gene disruption, andreintroduction of the deleted gene back to the mutant. Allthe introduced exogenous DNAs in the gene disruptionprocedures were removed from the final mutant strainexcept the two 34 bp loxP pieces left in deleted gene loci.
Ning, Fangkun,Wang, Chao,Berry, Karin Zemski,Kandasamy, Pitchaimani,Liu, Haolin,Murphy, Robert C.,Voelker, Dennis R.,Nho, Chu Won,Pan, Choel-Ho,Dai, Shaodong,Niu, Liwen,Chu, Hong-Wei,Zhang, Gongyi The Federation of American Societies for Experimen 2014 The FASEB Journal Vol.28 No.12
<P>The short palate, lung and nasal epithelial clone 1 (SPLUNC1) protein is a member of the palate, lung, and nasal epithelium clone (PLUNC) family, also known as bactericidal/permeability-increasing (BPI) fold-containing protein, family A, member 1 (BPIFA1). SPLUNC1 is an abundant protein in human airways, but its function remains poorly understood. The lipid ligands of SPLUNC1 as well as other PLUNC family members are largely unknown, although some reports provide evidence that lipopolysaccharide (LPS) could be a lipid ligand. Unlike previous hypotheses, we found significant structural differences between SPLUNC1 and BPI. Recombinant SPLUNC1 produced in HEK 293 cells harbored several molecular species of sphingomyelin and phosphatidylcholine as its ligands. Significantly, <I>in vitro</I> lipid-binding studies failed to demonstrate interactions between SPLUNC1 and LPS, lipoteichoic acid, or polymyxin B. Instead, one of the major and most important pulmonary surfactant phospholipids, dipalmitoylphosphatidylcholine (DPPC), bound to SPLUNC1 with high affinity and specificity. We found that SPLUNC1 could be the first protein receptor for DPPC. These discoveries provide insight into the specific determinants governing the interaction between SPLUNC1 and lipids and also shed light on novel functions that SPLUNC1 and other PLUNC family members perform in host defense.—Ning, F., Wang, C., Berry, K. Z., Kandasamy, P., Liu, H., Murphy, R. C., Voelker, D. R., Nho, C. W., Pan, C.-H., Dai, S., Niu, L., Chu, H.-W., Zhang, G. Structural characterization of the pulmonary innate immune protein SPLUNC1 and identification of lipid ligands.</P>