一种新的近交系实验动物遗传监测方法及小鼠性别相关RAPD标记的发现

我们用21个10 bp的随机短引物对来自昆明、成都、上海、北京四个地方的BALB/c小鼠以及C57BL小鼠、昆明种小白鼠进行了随机扩增多态DNA(RAPD)分析,发现13个引物的扩增产物在BALB/c小鼠和C 57 BL小鼠中有差异,8个引物的扩增产物在BALB/c小鼠和昆明种小白鼠之间不同.在四个地方的BALB/c小鼠中,成都、上海、北京的BALB/c小鼠其遗传背景均一,而来自昆明的BALB/c小鼠中,有2只的4个引物的扩增产物不同于其它的BALB/c小鼠,表明这两只BALB/c小鼠可能曾发生过某种程度的遗传改变或污染。实验结果显示RAPD方法是一种有效的近交系实验动物遗传监测手段。实验中一个有趣的结果是,在OPG 2、OPE 4、OPE 9的扩增产物中,发现了严格的性别依赖的PAPD标记。OPE 9扩增产物中,凡雄性个体都有一条0.88 kb的标记.OPG 2、OPE 4则在所有的雄性个体中多扩增出一条约1.2 kb的带。通过交叉PCR扩增和斑点杂交证明OPG 2、OPE 4 得到的雄性特异性RAPD标记虽分子大小一致,但不具同源性。这些性别相关RAPD标记的染色体定位和性质分析正在进一步进行中.

A NEW WAY FOR INBRED STRAIN MICE GENETIC MONITORING AND THE DISCOVERY OF SEX-LINKAGING RAPD MARKERS

We used 21 10 bp random primers to amplify DNA for four colonies of BALB/ c mice, four individuals of C 57 BL mice, and four individuals of Kunming stock mice. The amplified band patterns were different between BALB/c and C 57 BL mice in the products of 13 primers, and 8 primer products showed difference between BALB/c and Kunming stock mice. These results indicated that we can easily distinguish different strains of mice by RAPD method. For the four colonies of BALB/c mice, the genetic background of Chengdu colony, Shanghai colony and Beijing colony were homogeneous at all RAPD markers, however, in Kunming colony, there were two BALB/c individuals were disclosed different amplified patterns by 4 primers, showing that these two Kunming BALB/c mice maybe suffer genetic contamination or mutation some time. An unexpected phenomenon discovered in this study is that sex specific amplification bands were amplified by primer OPG 2, OPE 4, OPE 9 in all male mice. Although an 1.2 kb band were disclosed by both OPG 2 and OPE 4 in male mice, cross RAPD and dot hybridization showed that the two bands were not homologous.