집누에나방 (Bombyx mori) chaperonin θ 유전자의 단리 및 발현 기구 = Cloning and Characterization of Chaperonin θ gene in the silkworm, Bombyx mori

다국어 초록 (Multilingual Abstract)

Chaperonin is protein complexes that assist the protein folding. These proteins belong to a large class of molecules that assist protein folding, called molecular chaperone. We isolated the chaperonin θ gene, in B. mori. Sequence analysis of B. mori chaperonin θ gene has 1931 bp of full-length cDNA. It contains an single ORF coding 545 amino acids. This gene detected a single copy in the genomic DNA and located in chromosome 2. In genome, chaperonin gene exist 6 exon/ 5 intron and about 5.5 kb lengths. The protein alignment with related homologous chaperonin has an overall 63~75% similarity to the silkworm. While there are differences between eukaryotic, bacteria and archaeal chaperonins the general structure are conserved. This gene transcripts were detected in early embryogenesis. It expressed up to hatching. Chaperonin θ mRNA would be detected in every tissue of larva (e.g, in fat body, mid-gut, gonads, silk glands and brain). These expression level continued through the post-developmental stage (pupa and moth). Notably, B. mori chaperonin θ gene was up-regulated by stress, bacteria and heat shock. Thus, it suggested that the function of chaperonin θ gene is related with the immune process and defense of stress.

목차 (Table of Contents)

• Ⅰ. 서론 = 1
• Ⅱ. 재료 및 방법 = 4
• 1. 공시재료 = 4
• 2. Chaperonin gene의 동정 = 4
• 3. 염기서열 상동성 분석 = 5
• 4. 유전자 구조 분석 = 5
• 4-1. Southern Hybridization = 5
• 5. 유전자의 발현분석 = 6
• 5-1. RNA 추출 = 6
• 5-2. Reverse Transcriptase PCR (RT-PCR) = 6
• 5-3. Real-time PCR = 6
• 5-4. Northern Hybridization = 7
• 6. 외부자극에 의한 유전자 발현 유도 분석 = 9
• 6-1. Heat shock = 9
• 6-2. Bacterial infection = 9
• Ⅲ. 결과 = 10
• 1. EST 분석에 의한 집누에나방 chaperonin θ (theta) 유전자의 발굴 = 10
• 2. 집누에나방 chaperonin θ 유전자의 특성 = 10
• 2-1. 집누에나방 chaperonin θ 유전자의 구조분석 = 10
• 2-2. 집누에나방 chaperonin θ 유전자의 발현 profiling = 18
• 2-2-1. 배발생기의 발현 pattern = 18
• 2-2-2. 발육 시기에 따른 발현 pattern = 18
• 3. 외부자극에 의한 chaperonin θ gene의 발현 profile = 28
• 3-1. Heat shock에 의한 발현 유도 = 28
• 3-2. Bacteria 감염에 의한 발현 유도 = 31
• Ⅳ. 고찰 = 35
• Ⅴ. 적요 = 38
• Ⅵ. 참고 문헌 = 39