The genetic polymorphism of swine leucocyte antigen (SLA) class I and class II region in the Lee-Sung strain of small-ear miniature pig

• Main Authors: Wu, Ming-Jen, 吳明真
• Other Authors: Sung Yung-Yi
• Format: Others
• Language: zh-TW
• Published: 1997
• Online Access: http://ndltd.ncl.edu.tw/handle/35556824571623822246

碩士 === 國立臺灣大學 === 畜產學系 === 85 === This study aimed at the genetic polymorphism of swine leucocyte antigen(SLA) class I and class II region in the Lee- Sung strain of small-ear miniaturepig for researching about the SLA genotype distribution in this population andsupporting a good animal model in transplantation immunology research. Bloodsample collects from 84 Lee-Sung strain of small-ear miniature pigs, 5 small-earminiature pigs, and one male and one female Landrace. Also, there are 47 DNAsamples of small-ear miniature pig from animal germplasm conservation . Afterisolating DNA from buffy coat with isolation kit, using optimal primer to runpolymerase chain reaction (PCR) to obtaining the target product, and checkingit with few product. Others divide to equal part to restriction fragment lengthpolymorphism (RFLP) analysis. The SLA-DQB region digests by restriction enzymeHae III and Rsa I and SLA-DRB region by Msp I and Rsa I. The result shows that :(1) After using the primer 1 and 5, the PCR product in the SLA class I region isabout 210bp, but not good in specificity and repetition within this population.(2) In the SLA class II region, use the primer SgDQB5 a/SgDQB3a, SgDQB5a/SgDQB3bin SLA-DQB loci and the primer SgDRB5 a/ SgDRB3a, SgDRB5a/SgDRB3b in SLA-DRB loci, the average PCR product length are about 280bp, 270bp in SLA-DQB loci andabout 265bp, 245bp in SLA-DRB loci. After RFLP analysis, the SLA-DQB loci isputative HA type by Hae III digested and RA type by Rsa I digested. They arepossible recombination of e, g type by Hae III digested and c, g mixed type byRsa I digested, compared to the literature of Shia (1995). Also in the SLA-DRBloci, the genotype is putative MA type by Msp I digested and RA type by Rsa Idigested. In comparison with the literature of Shia (1995), they are possible atype by Msp I digested and b type by Rsa I digested. But in the DNA sample ofsmall-ear miniature pig from animal germplasm conservation, the SLA-DQB PCR-RFLPpatterns have three putative type by Hae III digested (HA, HB, and HC) and fourputative type by Rsa I digested (RA, RB, RC, and RD). In comparison with theliterature of Shia (1995), they are possible recombination of e, g type (HA), d,e mixed type (HB) and e, g mixed type (HC) by Hae III digested and c, g mixedtype (RA), a, c, g mixed type (RB), c type (RC) and g type (RD) by Rsa Idigested. This result is not consistent with the Lee-Sung strain of small-earminiature pig, means that there if no genetic polymorphism of SLA region in thispopulation after long-term inbred breeding, but the real serological type andfull sequence need further to be studied.