초록 ▼

1. 항 미생물 펩타이드(antimicrobia1 peptide)란 무엇인가?
생명과학 분야에 종사하고있는 많은 연구자들은 자연에는 현재 인류가 파악하고 있지 못한 질병치료에 대한 열쇠가 숨겨져 있다고 믿고 있다. 이는 동양의학에서의 한약재료로서 혹은 우리나라의 전통적인 민간요법으로서 생물체를 직접 복용한다는 사실로써도 입증되는 것이라고 볼 수 있다. 그러므로 최근 전 세계적으로 활발하게 진행되고았는 바, 인류가 겪고 있는 질병, 특히 감염성 질병(infectious disease)에 대한 치료제를 생물체로부터 얻으려 하는 연

1. 항 미생물 펩타이드(antimicrobia1 peptide)란 무엇인가?
생명과학 분야에 종사하고있는 많은 연구자들은 자연에는 현재 인류가 파악하고 있지 못한 질병치료에 대한 열쇠가 숨겨져 있다고 믿고 있다. 이는 동양의학에서의 한약재료로서 혹은 우리나라의 전통적인 민간요법으로서 생물체를 직접 복용한다는 사실로써도 입증되는 것이라고 볼 수 있다. 그러므로 최근 전 세계적으로 활발하게 진행되고았는 바, 인류가 겪고 있는 질병, 특히 감염성 질병(infectious disease)에 대한 치료제를 생물체로부터 얻으려 하는 연구는 새로운 type의 항쟁제를 고안해내기 위한 매우 가치 있는 시도라고 보겠다.

Abstract ▼

An insect defensin (Galleria defensin) was purified from the larval hemolymph of Galleria mellonella immunized against E. coli. The peptide composed of 43 amino acid residues containing 6 cysteines might be engaged in intramolecular disulfide bridges. The primary structure of Galleria defensin share

An insect defensin (Galleria defensin) was purified from the larval hemolymph of Galleria mellonella immunized against E. coli. The peptide composed of 43 amino acid residues containing 6 cysteines might be engaged in intramolecular disulfide bridges. The primary structure of Galleria defensin shared about 90.7% identity to that of heliomicin, which is an insect defensin isolated from Heliothis virescens. The full-length cDNA encoding Galleria defensin was cloned from the fat body of the immunized G. mellonella larvae. Northern blot analysis revealed that Galleria defensin was expressed not only in the fat body but also in the midgut against invading bacteria into hemocoel. Cecropin-like antimicrobial peptide, named Gm cecropin, was purified from hemolymph of the immunized wax moth, Galleria mellonella larvae, against E. coli and the antibacterial activity of Gm cecropin was examined in radial diffusion assay. The molecular mass of Gm cecropin was measured to be 4,160.69 Da in matrix-assisted laser desorption ionization-time-of-flight mass spectrometry analysis. Also, the full-length cDNA of the Gm cecropin precursor was cloned by a combination of RT-PCR using a designed degenerate primer that was originated for the N-terminal sequence by Edman degradation and 5-RACE-PCR. The cDNA structure of Gm cecropin showed that it was synthesized as a prepropeptide, which contains a putative 22-residue signal peptide, a 4-residue propeptide and followed by 39-residue mature peptide with calculated mass of 4,344.18 Da. From difference between the calculated and measured mass values, Gm cecropin was surmised to be a 37-residue peptide generated by the truncation of C-terminal residue and the amidation process.
Lysozymes were purified from the hemolymph of three immunized Lepidopteran larvae, Gallriamellonella, Bombyx mori, Agrius convolvuli to compare their physico-chemical properties and antibacterial activities with those of chicken lysozyme. Four lysozymes including chicken one had a similar molecular masses and chromatographic behavior on RP-HPLC. In Western blotting analysis, antibody raised against G. mellonella lysozyme was cross-reacted with other two insect lysozymes but not with the commercial chicken lysozyme. Antibacterial activities of lysozymes were measured in two types of tests: radial diffusion assay and colony count assay. Our antibacterial tests revealed that all lysozymes have strong activities against Gram-positive bacteria and three insect lysozymes still retain a little potency against Gram-negative bacteria, while chicken lysozyme has no activity against Gram-negative bacteria. Taken together, three Lepidopteran lysozymes have a common distinct str ucture to act on Gram-negative bacteria.

목차 Contents

• 표지 ...1
• 제출문 ...4
• 목차 ...6
• 제1세부과제명 : 약용곤충유래 항진균 물질의 탐색 및 이용.제2세부과제명 : 약용곤충으로부터 항균 펩타이드 분리 및 응용 ...8
• 영문요약(Summary) ...10
• I. 서론 ...12
• II. 연구사 ...17
• III. 재료 및 방법 ...24
• IV. 결과 및 고찰 ...31
• V. 적요 ...44
• VI. 인용문헌 ...46
• 제3 세부과제명 : 약용 곤충의 면역활성능의 탐색과 이용기술 개발 ...50
• 영문요약(Summary) ...52
• I. 서론 ...55
• II. 연구사 ...57
• III. 재료 및 방법 ...63
• IV. 결과 및 고찰 ...69
• V. 적요 ...95
• VI. 인용문헌 ...98