표제지
목차
Abstract 9
Ⅰ. 서론 10
Ⅱ. 재료 및 방법 13
1. Next Generation Sequencing (NGS) 분석 13
2. 분자적 특성 13
3. 유전자 발현분석 14
1) 정상 어체에서의 조직별 발현분석 14
2) 병원체 인위감염 후 시간별 발현분석 16
4. 항균펩타이드의 항균 활성 17
1) 펩타이드 합성 17
2) 균주 18
3) Minimum inhibitory concentration (MIC) 18
4) 세포 독성 18
5) Biofilm assay 19
Ⅲ. 결과 20
1. 분자적 특성 20
1) Sequences 분석 20
2) Multiple alignment 분석 22
3) 계통발생학적 분석 24
2. 유전자 발현분석 26
1) 정상 어체 조직별 발현분석 26
2) 병원체 인위감염 후 시간별 발현분석 28
3. 항균 펩타이드의 항균 활성 30
1) Minimum inhibitory concentration (MIC) 30
2) 세포 독성 31
3) Biofilm assay 33
Ⅳ. 고찰 35
Ⅴ. 요약 38
Ⅵ. 참고문헌 39
Table 1. PCR primers used in this study 15
Table 2. Synthesis peptides used in this study 17
Table 3. Antimicrobial activity of RPS27A and S30FAU. 30
Fig. 1. cDNA and deduced amino acid sequence of the RPS27A (A), S30FAU (B). The box indicate UBQ and Ribosomal_S27 domain in (A), box... 21
Fig. 2. Multiple sequence alignment analysis of starry flounder RPS27A (A) and S30FAU (B) sequence with other species amino acid sequences. NCBI... 23
Fig. 3. Phylogenetic analysis of deduced RPS27A (A) and S30FAU (B) amino acid sequences in fish. The phylogenetic tree was constructed using... 25
Fig. 4. Detection of RPS27A (A), S30FAU (B) genes in different tissues of healthy starry flounder by real-time PCR. EF-1α was used for normalizing... 27
Fig. 5. Gene expression of RPS27A (A) and S30FAU (B) in the kidney, spleen, gill, liver, brain, intestine, heart after infection with VHSV and S.... 29
Fig. 6. The haemolytic and cytotoxicity of RPS27A (A), S30FAU (B) were analysed using starry flounder red blood cells. The positive control was... 32
Fig. 7. Inhibitory effect of RPS27A (A), S30FAU (B) on bacterial biofilm formation. Values are expressed as the mean ± SD, and asterisks indicate a... 34