摘要
目的 选育猪到人异种移植的供器官猪种和医药实验研究大动物用猪种 ,首先应筛查其是否易患猪应激综合征 (PSS)。方法 采用聚合酶链反应结合限制性内切酶技术 (PCR- RFL P)研究版纳小耳猪近交系(BMI)和近交培育之中的五指山猪 (WZSP)的氟烷基因型 ,筛查其是否有易患猪应激综合征的可能。结果 5 0头版纳小耳猪近交系和 15头五指山猪氟烷基因型均为阴性纯合子。结论 这两种猪种可望成为猪到人异种移植和医药实验研究的理想动物。
Objective Porcine stress syndrome (PSS) is one kind of molecular genetics defect diseases of pig which will cause malignant hyperthermia syndrome(MHS) and is the first index should be excluded in screening of a pig species for xenotransplantation. It was reported that mutation of pig rynodine receptor(RYR1) gene is the main reason for PSS. In this study, RYR1 genotypes of the Chinese Banna mini pig inbred line and inbreeding closed colony Wuzhishan pig were investigated with polymerase chain reaction-restriction endonuclease fragment length polymophism(PCR-RFLP) technique. Methods Antevenocaval whole blood samples were collected from 50 Banna mini-pig inbred-line(BMI), 15 inbreeding Wuzhishan pig (WZSP) and 25 Neijiang pigs(NJP) as negative control, the primer were designed and synthesized, PCR reaction was conducted following the sequence of 94℃(1 min), 58℃(1 min) and 72℃(1 min) for 30 cycles. The PCR products were digested with restriction endonuclease HhaI and then electrophoresis check. Results A 659 bp DNA fragment was amplified with these two primers,the HAL NN sample fragment was cut into fragments as 493 bp and 166 bp individually after the digestion, indicates no point mutation at site 1 843 in RYR1 gene in all tested BMI pig and WZSP. Namely, the RYR1 genotype of 50 cases of BMI and 15 cases of WZSP were HAL NN , therefore their phenotype is PSS negative. Conclusion It indicates that the genotype of Banna mini pig inbred line and inbreeding Wuzhishan pig are HAL NN therefore PSS absolutely negative, the group penetrance is 0. This is consistent with experimental observation. It suggestes that Banna mini pig inbred line and inbreeding Wuzhishan pig may be the alternative donor for xenotransplantation.
出处
《中国修复重建外科杂志》
CAS
CSCD
2000年第5期311-314,共4页
Chinese Journal of Reparative and Reconstructive Surgery
基金
国家自然科学基金!重大项目资助 (39993430 )
关键词
近交系猪
氟烷基因型
异种移植
PCR-RFLP
Inbred-line pig Rynodine receptor genotype Polymerase chain reaction-restriction endonuclease fragment length polymophism Xenotransplantation
