【作者】 周波；

【导师】 王林云； 张似青；

【作者基本信息】 南京农业大学 ， 动物遗传育种与繁殖， 2003， 硕士

【摘要】 采用PCR-RFLP法对中国地方猪种五指山、二花脸及国外猪种皮特兰猪的SLA-DQB和DRB基因外显子2的PCR产物进行了分析。SLA-DQB基因外显子2经限制酶HaeⅢ(7种RFLP带型)和Rsal(6种RFLP带型)酶切后共分出9种RFLP带型，五指山、二花脸和皮特兰猪分别分出3、3、5种RFLP带型；SLA-DRB基因外显子2经限制酶MspⅠ(1种RFLP带型)和RsaⅠ(5种RFLP带型)酶切后共分出5种RFLP带型，五指山、二花脸和皮特兰猪分别分出2、3、3种RFLP带型。 通过RFLP带型分析后，将带型为AA的五指山猪的SLA-DQB基因外显子2的PCR产物进行克隆测序，结果发现了一个新的等位基因SLA-DQB*NAU。DNA序列已经提交美国国家生物信息技术中心(NCBI)，登录号为AY281361。这一新SLA-DQB等位基因在抗原肽结合槽处发生了一个氨基酸突变。 将组合RFLP带型BCFGHHIJ标记为a带型，组合RFLP带型CCFFHHII标记为b带型。对a、b两种带型的二花脸猪进行血清IgG含量、淋巴细胞转化率和PHA皮肤试验等抗病力指标的检测，来分析不同RFLP带型猪之间差异。结果表明：基因型为杂合子(a带型)的猪比纯合子(b带型)的猪在对抗疾病的免疫能力上有一定的差异，杂合子更具优势。更多还原

【Abstract】 College of Animal Science and Technology, Nanjing Agricultural University, 210095, P. R. ChinaThe restriction polymorphism of the exon 2 of SLA-DQB and DRB gene in Wuzhishan, Erhualian and Pietrain Pigs was investigated by PCR-RFLP. Nine kinds of SLA-DQB PCR-restriction fragment length polymorphism (RFLP) pattern types were observed for both HaeIII (7 RFLP pattern types) and RsaI(6 RFLP pattern types). There were 3, 3 and 5 kinds of PCR-RFLP pattern types in Wuzhishan, Erhualian and Pietrain Pigs, respectively. A total of five kinds of SLA-DRB PCR-RFLP pattern types were defined using MspI (1 RFLP pattern types) and Rsal (5 RFLP pattern types). There were 2, 3 and 3 kinds of PCR-RFLP pattern types in Wuzhishan, Erhualian and Pietrain Pigs, respectively.The PCR product of PCR-RFLP pattern type of A in Wuzhishan was cloned into vector and sequenced using standard protocols, and an automated sequencer. As a result, a new SLA-DQB allele was found in Wuzhishan, a Chinese native pig. The DNA sequence has been submitted to the Genbank DNA Sequence Database under accession number AY281361. This new allele sequence has a new mutation in the peptide-binding groove.The PCR-RFLP pattern type of BCFGHHIJ in both SLA-DQB and DRB gene was named a pattern type, and the PCR-RFLP pattern type of CCFFHHII in both SLA-DQB and DRB was named b pattern type. Three immunological indexes, such as the transformation ratio of lymphocyte, the content of IgG in serum and the PHA test of skin, were measured in the pigs of the two PCR-RFLP pattern types. As a result, the abilitiy of pigs of heterozygote (a pattern type) was stronger than it of homozygote (b pattern type) on disease resistance.更多还原