Sanyal A, O'Driscoll S W, Fitzsimmons J S, Bolander M E, Sarkar G
Cartilage and Connective Tissue Research Laboratory, Mayo Clinic, Rochester, MN 55905, USA.
Mol Biotechnol. 1998 Dec;10(3):223-30. doi: 10.1007/BF02740842.
A method termed "gene digging" has been developed based on our observation of stretches of highly conserved nucleotide sequence in the coding region of many genes across related species. Rabbit-specific nucleotide sequences corresponding to desired coding segments of 14 different genes were obtained with primers that were designed based on conserved nucleotide stretches. Our success in gene digging could be attributable to the method's inherent ability to reduce the degeneracy of primers by more than two orders of magnitude (sometimes by more than three orders of magnitude) compared to primers designed from conserved amino acids. Our results not only demonstrate the value of the method, but also hint at a thus far unknown functional significance of conserved nucleotide stretches in the coding region of various genes. In our hands the method worked 14 out of 14 times indicating generality of the concept.
基于我们对相关物种中许多基因编码区高度保守核苷酸序列片段的观察，已开发出一种名为“基因挖掘”的方法。利用基于保守核苷酸片段设计的引物，获得了与14个不同基因的所需编码片段相对应的兔特异性核苷酸序列。与根据保守氨基酸设计的引物相比，我们在基因挖掘方面的成功可能归因于该方法固有的将引物简并度降低两个以上数量级（有时超过三个数量级）的能力。我们的结果不仅证明了该方法的价值，还暗示了各种基因编码区中保守核苷酸片段迄今未知的功能意义。在我们的操作中，该方法在14次实验中有14次成功，表明了这一概念的通用性。
