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이종이식에 활용할 α1,3-galactosyltransferase 비활성화 및 Membrane Cofactor Protein 발현 동형접합 형질전환 돼지 개발
이건섭,박상현,이해선,지수정,이주영,변승준,황성수,김경운,옥선아,오건봉,Lee, Gunsup,Park, Sang Hyoun,Lee, Haesun,Ji, Soo-Jeong,Lee, Joo Yung,Byun, Sung-June,Hwang, Seongsoo,Kim, Kyung Woon,Ock, Sun A,Oh, Keon Bong 한국수정란이식학회 2017 한국동물생명공학회지 Vol.32 No.3
Transplantation is considered to be a very useful approach to improve human welfare and to prolong life-span. Heterologous organ transplantation using pig organs which are similar to human beings and easy to make mass-production has known as one of the alternatives. To ensure potential usage of the pig organ for transplantation application, it is essentially required to generate transgenic pig modifying immuno-related genes. Previously, we reported production of heterozygous ${\alpha}1,3$-galactosyltransferase (GalT) knock-out and human membrane cofactor protein (MCP) expressing pig ($GalT^{-MCP/+}$), which is enforced for suppression of hyperacute and acute immunological rejection. In this study, we reported generation of homozygous pig ($GalT^{-MCP/-MCP}$) by crossbreeding $GalT^{-MCP/+}$ pigs. Two female founders gave birth to six of $GalT^{-MCP/-MCP}$, and seven $GalT^{-MCP/+}$ pigs. We performed quantitative real-time PCR, western blot, and flow cytometry analyses to confirm GalT and MCP expression. We showed that fibroblasts of the $GalT^{-MCP/-MCP}$ pig do not express GalT and its product Gal antigen, while efficiently express MCP. We also showed no expression of GalT, otherwise expression of MCP at heart, kidney, liver and pancreas of transgenic pig. Taken together, we suggest that the $GalT^{-MCP/-MCP}$ pig is a useful candidate to apply xenotransplantation study.
3D8 scFv 형질전환 돼지 개발 및 PRRS 저항성 평가
이휘철 ( Hwi-cheul Lee ),이건섭 ( Gunsup Lee ),김지윤 ( Ji-yoon Kim ),양현 ( Hyeon Yang ),이보람 ( Bo Ram Lee ),박미령 ( Mi-ryung Park ),황인설 ( In-sul Hwang ),이풍연 ( Poongyeon Lee ),변승준 ( Sung-june Byun ),김원일 ( Won-il K 한국동물위생학회(구 한국가축위생학회) 2020 韓國家畜衛生學會誌 Vol.43 No.4
In this study, we have developed 3D8 scFv transgenic pig (TG) by microinjection of fertilized one-cell pig zygotes (2.17%). The effect of 3D8 scFv TG on porcine reproductive and respiratory syndrome virus (PRRSV) resistance were evaluated through PRRSV VR2332 (1×10 <sup>3</sup> TCID<sub>50</sub>/mL) challenge and transmission experiments. As a result, the average daily weight gain (ADWG) of TG increased compared to the wild type pigs (WT) in PRRSV challenge groups and the serum viremia levels of the TG was significantly lower than of WT on the 7 day and 21 day after infection, meaning that the viral shedding was suppressed by 3D8 scFv expression. These results suggest that the expression of 3D8 scFv in pig could suppress spreading of infected virus to pigs sharing a room.