A high frequency of Mamu-A*01 in the rhesus macaque detected by polymerase chain reaction with sequence-specific primers and direct sequencing
A high frequency of Mamu-A*01 in the rhesus macaque detected by polymerase chain reaction with sequence-specific primers and direct sequencing
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DOI：
10.1111/j.1399-0039.1997.tb02927.x
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发表时间：
1997-12-01
期刊：
影响因子：
--
通讯作者：
Watkins, DI
中科院分区：
文献类型：
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作者：
Knapp, LA;Lehmann, E;Watkins, DI
SIV感染恒河猴是HIV感染人类的一个很好的模型。不幸的是，很难鉴定表达特定MHC I类等位基因的猕猴。在这里，我们描述了使用PCR-SSP的恒河猴Mamu-A*01分型。使用Mamu-A * 01特异性引物从基因组DNA中扩增Mamu-A *01等位基因，并将阳性PCR产物直接测序。我们的技术在68只受试动物中鉴定出15只Mamu-A*01阳性动物。我们验证了我们的分子分析表明，从8个Mamu-A*01阳性动物的淋巴细胞表达Mamu-A*01确定的免疫沉淀和1-D IEE技术的简单性和准确性，这种分型方法应有利于选择Mamu-A*01阳性恒河猴艾滋病病毒的发病机制和疫苗的研究。
SIV infection of rhesus macaques is an excellent model for HIV infection of humans. Unfortunately, it is has been difficult to identify macaques expressing particular MHC class I alleles. Here we describe the use of PCR-SSP for Mamu-A*01 typing of rhesus macaques. The Mamu-A*01 allele was amplified from genomic DNA using Mamu-A*01-specific primers and positive PCR products were directly sequenced. Our technique identified 15 Mamu-A*01-positive animals of 68 tested. We validated our molecular analysis by showing that lymphocytes from 8 Mamu-A*01-positive animals expressed Mamu-A*01 as determined by immunoprecipitation and 1-D IEE The technical simplicity and accuracy of this typing method should facilitate selection of Mamu-A*01-positive rhesus macaques for AIDS virus pathogenesis and vaccine studies.