아주대학교

목차

1. Introduction 1
2. Materials and Methods 5
2.1 Strains and media 5
2.2 Plasmid construction 5
2.3 Site-directed mutagenesis 6
2.4 Strain construction 7
2.5 Shake flask cultivation 7
2.6 Transcriptional analysis (RT-PCR) 8
2.7 Extraction of C20-, C25- and C30-phytoene 8
2.8 Analysis of HPLC and mass spectrometry (MS) 9
3. Results 16
3.1 Construction and expression analysis of genes for an alternative carotenoid biosynthetic pathway 16
3.2 Integration of CrtMLIKE (C30-phytoene synthase) gene in S. cerevisiae 18
3.3 Assessment of CrtMLIKE expression efficiency in plasmid-based and genome-integrated systems 21
3.4 Engineering a GPP-accumulation platform strain through site-directed mutagenesis of ERG20 24
3.5 Kinetic analysis of 4,4'-diapophytoene production in recombinant S. cerevisiae 27
3.6 Codon optimization of CrtMLIKE to improve expression in S. cerevisiae 29
3.7 Construction of a C20, 25-phytoene biosynthetic pathway in S. cerevisiae 31
4. Discussion 33
5. References 36
ABSTRACT IN KOREAN 40