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소 β-casein 유전자 영역에서 소 Insulin-like Growth Factor 1을 생산하기 위한 Knock-in Vector
김상영,박다솜,김세은,구덕본,강만종,Kim, Sang Young,Park, Da Som,Kim, Se Eun,Koo, Deog-Bon,Kang, Man-Jong 한국동물번식학회 2017 Reproductive & developmental biology Vol.41 No.3
The production of therapeutic protein from transgenic domestic animal is the major technology of biotechnology. Insulin-like growth factor-1 (IGF-1) is known to play an important role in the growth of the animal. The objective of this study is construction of knock-in vector that bovine IGF-1 gene is inserted into the exon 7 locus of ${\beta}$-casein gene and expressed using the gene regulatory DNA sequence of bovine ${\beta}$-casein gene. The knock-in vector consists of 5' arm region (1.02 kb), bIGF-1 cDNA, CMV-EGFP, and 3' arm region (1.81 kb). To express bIGF-1 gene as transgene, the F2A sequence was fused to the 5' terminal of bIGF-1 gene and inserted into exon 7 of the ${\beta}$-casein gene. As a result, the knock-in vector is confirmed that the amino acids are synthesized without termination from the ${\beta}$-casein exon 7 region to the bIGF-1 gene by DNA sequence. These knock-in vectors may help to create transgenic dairy cattle expressing bovine bIGF-1 protein in the mammary gland via the expression system of the bovine ${\beta}$-casein gene.
돼지 $\beta$-Casein을 이용한 EGFP 발현 Knock-in 벡터의 구축 및 발현 검증
이상미,김혜민,문승주,강만종,Lee, Sang-Mi,Kim, Hey-Min,Moon, Seung-Ju,Kang, Man-Jong 한국동물번식학회 2008 Reproductive & developmental biology Vol.32 No.3
This study was carried out to develop knock-in vector for EGFP (enhanced green fluorescent protein) expression in porcine $\beta$-casein locus. For construction of knock-in vector using porcine $\beta$-casein gene, we cloned the $\beta$-casein genome DNA from porcine fetal fibroblast cells, EGFP and SV40 polyA signal using PCR. The knock-in vectors consisted of a 5-kb fragment as the 5' recombination arm and a 2.7-kb fragment as the 3' recombination arm. We used the neomycin resistance gene ($neo^{r}$) as a positive selectable marker and the diphtheria toxin A (DT-A) gene as a negative selectable marker. To demonstrate EGFP expression from knock-in vector, we are transfected knock-in vector that has EGFP gene in murine mammary epithelial cell line HC11 cells with pSV2 neo plasmid. The EGFP expression was detected in HC11 cells transfected knock-in vector. This result demonstrates that this knock-in vector may be used for the development of knock-in transgenic pig. 본 연구는 돼지 $\beta$-casein 유전자 위치에서 EGFP가 발현될 수 있는 knock-in 벡터를 구축하기 위하여 실시되었다. 돼지의 $\beta$-casein 유전자를 이용하여 knock-in 벡터를 구축하기 위해 돼지의 태아 섬유아세포로부터 $\beta$-casein 유전자를 동정하였고 EGFP, SV4O polyA signal을 동정하였다. Knock-in 벡터는 5' 상동 영역 약 5 kb와 3' 상동 영역 약 2.7 kb로 구성되어있으며, positive selection marker로 $neo^{r}$ 유전자를, negative selection marker로 DT-A 유전자를 사용하였다. 구축된 knock-in 벡터로부터 EGFP의 발현을 확인하기 위하여 생쥐 유선 세포인 HC11 세포에 knock-in 벡터를 도입하였다. 그 결과 EGFP의 발현을 HC11 세포에서 확인하였다. 이와 같은 결과로서 이 block-in 벡터는 knock-in 형질전환 돼지를 생산하는데 사용될 수 있을 것으로 생각된다.
프로테아제, 자일라나제, 만나아제의 첨가가 비육돈의 성장, 혈액성상 및 생산성에 미치는 영향
고대건(Dae-Geon Go),이용기(Yong-Gi Lee),유기명(Ki-Myeong Yoo),선현수(Hyen-Su Seon),윤진혁(Jin-Hyeok Yoon),윤상(Sang Yoon),김승연(Seung-Yeon Kim),유형주(Hyeong-Ju You),김민석(Min-Seok Kim),이지웅(Ji-Woong Lee),강만종(Man-Jong Kang) 한국산학기술학회 2021 한국산학기술학회논문지 Vol.22 No.10
본 연구에서는 비육돈 기초사료에 Protease, Xylanase 및 Mannanase를 포함한 복합효소제를 첨가하였을 때 육성돈의 성장 및 경제성에 미치는 영향에 대하여 조사하기 위하여 실시하였다. 실험을 위하여 평균체중 37.48 ± 5.43kg인 비육돈(Landrace × Yorkshire × Duroc) 1478 두를 공시하여 3반복하고, 반복 당 대조구는 평균 256 두, 처리구는 평균 236 두씩 완전임의 배치하였다. 처리구는 Protease + Xylanase + Mannanase 0.1 % 처리하여 출하체중 약 115 kg까지 사양실험을 실시하였다. 사양성적에 있어 일당증체량은 처리구와 대조구에서 각각 784.62 ± 17.42와 대조구 749.39 ± 10.48로 처리구에서 유의적으로 개선되었다(p >0.05). 사료섭취량에서는 처리구간 유의차는 없었지만 (p >0.05) 사료요구율에서 대조구 0.355 ± 0.01, 처리구 0.365 ± 0.01로 처리구에서 유의적으로 개선되었다(p <0.05). 혈액 성상에서는 Protease, Xylanase 및 Mannanase 첨가구가 대조구에 비해 Total protein 함량이 유의적으로 높게 나타났다(p <0.05). 경제성 분석에서는 두당 증체량(total weight gain)과 1 두당 돈가(total pig prices)의 경우 처리구에서 대조구보다 개선되는 경향을 보였지만 유의적 차이는 없었다(p >0.05). 1등급 이상 돼지 출현율(Prevalence of grade 1 or higher)은 처리구와 대조구에서 각각 57.41 ± 0.4, 49.54 ± 4.92로 처리구에서 유의적으로 개선되었다(p <0.05). 본 실험의 결과로 보아 Protease, Xylanase 및 Mannanase를 포함한 복합효소제를 비육돈 기초사료에 첨가하는 것은 돼지의 성장을 촉진하여 생산성 및 경제성이 개선하고 혈액 내 단백질함량을 증가시키는 것으로 나타난다. This study was conducted to investigate the effect of supplementation with an enzyme complex including protease, xylanase, and mannanase to the basal diets of pigs on their growth performance and economic value. A total of 1478 cross-bred-fed pigs (Landrace × Yorkshire × Duroc) of 37.48 ± 5.43 kg (average BW) were randomly assigned to one of these treatment groups with 3 repetitions. Breeding experiments of the group supplemented with 0.1 % enzyme complex were conducted by attaining a bodyweight of about 115 kg. Growth performance improved significantly with supplementation treatment compared to the basal diet showing an average daily gain of 749.39 ± 10.48 and 784.62 ± 17. 42 (p >0.05). The feed conversion ratio significantly improved in the supplemented groups at 0.365 ± 0.01 compared with 0.355 ± 0.01 in non-supplemented groups (p <0.05). An analysis of blood composition showed that the total protein in the treatment group was significantly higher than the control (p <0.05). Total weight gain and
강만종,이철상,한용만,유대열,이경광 제주대학교 농과대학 제주도축산문제연구소 1991 畜産論叢 Vol.6 No.1
This study was carried out in order to investigate eggects of cryoprotectant concentration and equilibration time on survival of ultrarapidly frozen 2-cell mouse embryos Mouse 2-cell embryos. fol-lowing dehydration by exposure to DMSO and sucrose. were directly immersed into liquid nitrogen and thawed in 37℃ water. Viability was defined by development rate to the blastocyst stage after in vitro culture for 72 hours. The results are summarized as follows ; 1. When 0.25M of sucrose was added into the freezing medium a t various concentrations of DMSO and dilution medium, higher development rate of embryo was obtained in 3.0M DMSO conentrations (82.6%). However, When sucrose concentrations of 0.25 and 0.5M were added to the freezing medium with 3.0M DMSO and dilution medium, development rate of embryos were 81.7% and 24.1%, respectively. 2. In the equilibration time at room temperature, higher development rate was attained after short period of time(2.5min) in 3.0M DMSO +0.25M sucrose(85.9%). 3. The development rate of embryos at in vitro 2-celL in vivo 2-celL solution control and untreated control was 84.6%, 90.9%, 89.9%. and 89.7%. respectively.