Ko M S, Wang X, Horton J H, Hagen M D, Takahashi N, Maezaki Y, Nadeau J H
Center for Molecular Biology, Wayne State University, Detroit, Michigan 48202.
Mamm Genome. 1994 Jun;5(6):349-55. doi: 10.1007/BF00356553.
We recently proposed a new PCR-based genetic marker assay for the mouse genome that exploits sequence differences in the 3'-untranslated region (UTR) of cDNAs between different mouse strains, called "biallelic polymorphic expressed sequence tags (bESTs)." The specific use of 3'-UTR has several advantages: (1) frequent sequence polymorphism between different mouse strains, (2) most commonly uninterrupted by introns, (3) usually unique sequence even among closely related gene family members. In this paper, we identify additional genetic loci defined by bEST and determine their location on the mouse genetic map by using interspecific backcross mapping panels between C57BL/6J and Mus spretus. Of 136 markers tested, 86 produced unique PCR products from C57BL/6J and M. spretus genomic DNAs. We then sequenced these 86 PCR products from C57BL/6J and M. spretus and found that 59 markers have sequence polymorphisms. Of these, we mapped 36 by restriction fragment length polymorphism (RFLP) of the PCR products and 4 by length polymorphism (LP) of the PCR products. We discuss the possibility of a large-scale application of this method for cDNA mapping.
我们最近提出了一种基于PCR的小鼠基因组新遗传标记检测方法，该方法利用不同小鼠品系之间cDNA 3'非翻译区（UTR）的序列差异，称为“双等位基因多态性表达序列标签（bESTs）”。3'UTR的特殊用途具有几个优点：（1）不同小鼠品系之间频繁出现序列多态性；（2）大多数情况下不被内含子打断；（3）即使在密切相关的基因家族成员中通常也是独特序列。在本文中，我们鉴定了由bEST定义的其他遗传位点，并通过使用C57BL/6J和小家鼠之间的种间回交定位面板确定它们在小鼠遗传图谱上的位置。在测试的136个标记中，有86个从C57BL/6J和小家鼠基因组DNA中产生了独特的PCR产物。然后我们对来自C57BL/6J和小家鼠的这86个PCR产物进行了测序，发现59个标记具有序列多态性。其中，我们通过PCR产物中的限制性片段长度多态性（RFLP）定位了36个，通过PCR产物中的长度多态性（LP）定位了4个。我们讨论了这种方法大规模应用于cDNA定位的可能性。
