• 농업과학연구
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• 제28권2호
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• pp.92-98
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• 2001

락토페리신은 다양한 생리활성을 나타내는 락토페린(약 80kD)에서 유래된 항균성펩타이드 분획물(5kD)이다. 마우스HC11유선상피세포에서 인체 락토페리신의 발현은 bovine beta-casein을 promotor로 하고 인체 락토페리신 cDNA를 삽입하여 제작한 pBL1-cin발현벡타를 이용하였다. 이 발현벡타를 이용하여 인체 락토페리신 발현여부를 RT-PCR, northern blot, dot blot분석을 통하여 확인하였다. pBL1-cin 발현백타를 HC11세포에 transfection 하여 얻은 RNA를 이용하여 RT-PCR를 한 결과 150bp의 크기로 확인되었고 Northern blot 분석결과는 약 2.3 kb의 크기로 확인되었다. 인체 락토페린 polyclonal항체를 이용하여 dot blot한 결과 인체 락토페리신이 분비됨을 확인하였다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.134.2-135
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• 2003

We examined the minimal amino acid sequence of bovine lactoferricin (Lfcin-B), a cationic peptide corresponding to residues 17-41 near the N-terminus of bovine lactoferrin, to induce apoptosis in THP-l human monocytic leukemic cells using synthetic peptides. A synthetic peptide (Lfc-17/29, amino acid sequence; FKCRRWQWRMKKL) which is consist of 13 amino acids near the N-terminus of Lfcin-B induced cell death in THP-1 cells in a dose-dependent manner, showing apparent apoptotic changes such as hypodiploid forms of genomic DNA and apoptotic DNA fragmentation. (omitted)

• 한국수정란이식학회지
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• 제17권2호
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• pp.109-115
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• 2002

The present study was conducted for the production of transgenic cloned cows those secrete human lactoferricin into milk by somatic cell nuclear transfer (NT). To estimate detrimental effects of gene transfection on transgenic cloned embryo production, development rates of NT embryos were compared between transfected and non-transfected cumulus and ear fibroblast cells. An expression plasmid for human lactofericin (pbeta-LFC) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human lactoferricin target gene into a pcDNA3 plasmid. Two bovine somatic cell lines (cumulus cell and ear fibroblast) were established and transfected with the expression plasmid using a liposomal transfection reagent, Fugene6 as a carrier. Cumulus cell and ear fibroblast were transfected at the passage of 2 to 4, trypsinized and GFP-expressing cells were randomly selected and used for somatic cell NT. Developmental competences (rates of fusion, cleavage, and blastocyst formation) in bovine transgenic somatic cell NT embryos reconstructed with non-transfectecd cells were significantly higher than those from transfected cells in cumulus cell and ear fibroblast (P＜0.05). This study indicated that transfection of done. cell has detrimental effect on embryo development in bovine transgenic NT.

• Journal of Animal Science and Technology
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• 제45권4호
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• pp.537-542
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• 2003

개시시 체중 7.63, 0.41kg, 3원 교잡종 이유자돈(25일령) 60두를 공시하여, 이유자돈 사료내 재조합 인간 락토페리신 컬처(RHLC ; recombinant human lactoferricin culture)의 급여가 성장, 영양소 소화율 및 혈장내 면역글로블린 G 농도에 미치는 영향을 평가하기 위하여 20일간 사양시험을 실시하였다. 시험설계는 1) 항생제 무첨가구(NC), 2) 항생제 첨가구(PC; NC 사료 + 0.1% chlortetracycline), 3) 항생제 무첨가구 사료에 RHLC를 0.3% 첨가한 처리구 (RHLC0.3; NC 사료 + 0.3% RHLC), 4) 항생제 무첨가구 사료에 RHLC를 0.5% 첨가한 처리구 (RHLC0.5; NC 사료 + 0.5% RHLC)로 4개 처리로 하였다. 총 사양시험 기간동안, 일당증체량에 있어서는 항생제 첨가구와 비교하여 RHLC의 첨가수준이 증가함에 따라 높아지는 경향을 보였으나, 표준오차가 크기 때문에 유의적인 차이는 보이지 않았다. 일당사료섭취량에 있어서는 PC 처리구와 비교하여 RHLC0.3 처리구가 유의적으로 높았으며(P<0.05), 사료효율에 있어서는 PC 처리구와 비교하여 RHLC0.5 처리구가 가장 좋은 것으로 나타났다(P<0.05). 영양소 소화율에 있어서는 NC 처리구와 비교하여 PC, RHLC0.3 및 RHLC0.5 처리구가 유의적으로 높게 평가되었으며(P<0.05), 혈장내 면역글로블린 G 농도는 처리구간에 유의적인 차이를 보이지 않았다. 결론적으로, 이유자돈에 있어 재조합 인간 락토페린신 컬처가 항생제를 대체할 가능성이 있는 것으로 사료된다.

• BMB Reports
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• 제29권6호
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• pp.545-548
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• 1996

CA(1-8)ME(1-12), the CA-ME hybrid peptide of the amino terminal segments of cecropin A (CA) and melittin (ME), has been reported to have a broad spectrum and improved potency without a hemolytic property. In order to obtain new synthetic peptides with powerful antibacterial activity without hemolytic activity, several hybrid peptides were designed from the sequences of CA, ME, magainin 2, bombinin and lactoferricin. All hybrid peptides were constructed to form an amphipathically basic-flexible-hydrophobic structure and synthesized by the solid phase method. Their hemolytic activities against human red blood cells and antibacterial activities against both Gram-positive and Gram-negative bacteria were detennined. CA(1-8)MA(1-12), CA(1-8)BO(1-12), MA(10-17)ME(1-12) and LF(20-29)ME(1-12) showed comparable activities with broad spectra against both Gram-positive and Gram-negative bacteria relative to CA(1-8)ME(1-12) but without hemolytic properties. These hybrid peptides, therefore, could be useful as model peptides to design a novel peptide with improved antibacterial activity and study on structure-activity relationships of antimicrobial peptides.