Jorgensen C B, Wintero A K, Yerle M, Fredholm M
Division of Animal Genetics, Department of Animal Science and Animal Health, The Royal Veterinary and Agricultural University,Bülowsvej 13, 1870 Frederiksberg C, Copenhagen, Denmark.
Mamm Genome. 1997 Jun;8(6):423-7. doi: 10.1007/s003359900460.
Complementary DNA sequences were selected from a resource of tentatively identified clones from a porcine small intestine cDNA library. Forty PCR primer pairs were designed to amplify 101-309 base pairs of the 3' untranslated region of the genes. The PCR conditions were optimized by altering both formamide and magnesium concentrations on samples of pig, mouse, and hamster DNA. Twenty primer pairs that, under stringent conditions, were pig-specific and amplified the expected fragments were chosen for regional assignment in a pig/rodent hybrid cell panel. Furthermore, 22 primer pairs were chosen to amplify DNA from the parental animals of the PiGMaP shared reference families in order to detect possible polymorphisms. Primer pairs that generated polymorphisms were used for genetic mapping. A total of 22 porcine expressed sequence tags (ESTs) were cytogenetically or genetically mapped by this approach. Twelve of the mapped ESTs could be added to the human-porcine comparative map.
互补DNA序列是从猪小肠cDNA文库中初步鉴定的克隆资源中选取的。设计了40对PCR引物，用于扩增基因3'非翻译区101 - 309个碱基对。通过改变猪、小鼠和仓鼠DNA样本中的甲酰胺和镁离子浓度来优化PCR条件。选择了20对在严格条件下具有猪特异性且能扩增出预期片段的引物，用于在猪/啮齿动物杂交细胞系中进行区域定位。此外，选择了22对引物来扩增PiGMaP共享参考家系的亲本动物的DNA，以检测可能的多态性。产生多态性的引物对用于遗传图谱构建。通过这种方法，总共对22个猪表达序列标签（EST）进行了细胞遗传学或遗传学定位。其中12个已定位的EST可以添加到人类 - 猪比较图谱中。
