야로위아 리폴리티카의 Yloch1과 Yloch1 Ylmnn4 돌연변이로부터 분비된 재조합 단백질의 N-결합형 당쇄구조분석에 관한 연구 : Structural analysis of N-linked oligosaccharides assembled on recombinant proteins secreted from Yarrowia lipolytica Yloch1 and Yloch1 Ylmnn4 mutants

목차 (Table of Contents)

• Contents
• 1. Introduction...........................................................................................1
• 2. Materials and Methods...........................................................................4
• 2.1 Strain and media ...................................................................................4
• 2.2 Plsmid..................................................................................................4
• 2.3 Construction of disruption cassette...........................................................5
• 2.4 Purification of endoglucanase I and lipase ................................................5
• 2.5 Oligosaccharide preparation....................................................................6
• 2.6 HPLC analysis of N-linked oligosaccharides.............................................6
• 3. Results.................................................................................................9
• 3.1 Isolation of Y. lipolytica OCH1 gene..........................................................9
• 3.2 Effect of YlOCH1 deletion on cell growth and N-glycosylation.....................14
• 3.3 YlOCH1 as a functional homolog of ScOCH1 encoding an initiating？？a1,6-mannosyltransferase.............................................................................................18
• 3.4 Structural analysis of N-linked oligosaccharides released from EGI and lipase 2 in Yloch1∆ and Yloch1∆ Ylmnn4∆................................................................................20
• 4. Disccussion.........................................................................................28
• 5. References...........................................................................................31
• 6. Abstract ..............................................................................................37