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Construction of the Rat Chromosomal Map by DNA Probes.

通过 DNA 探针构建大鼠染色体图谱。

基本信息

批准号：
01440088
负责人：
YAMADA Junzo
金额：
$ 4.42万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01440088/
关键词：
rat DNA probe situ hybridization gene mapping RFLP backcross analysis somatic cell hybridization in situ分子交雑法 DNAプロ-グ戻し交雑リンケ-ジ染色体in situ交雑法

项目摘要

1. Chromosomal assignment of the rat genes by DNA probes, backcross analysis, and in situ hybridization. (1) Angiotensinogen (Ang) : 3 alleles of Ang were detected among inbred strains of rats by using the cDNA and Southern blot analysis. Ang was not linked with other 16 loci using 2 sets of backcross analysis, but was assigned to 19q region by in situ hybridization. (2) Insulin-1 (Ins-1) : Ins-1 was assigned to 1q55 by fluorescence in situ hybridization (FISH) , and 4 alleles of the locus were detected by Southern blot analysis. Ins-1 was not inked with other 22 loci including c, Hbb which located on chromosome 1. This discrepancy may be due to the remote distance between them on the chromosome. (3) Renin (Ren) : Ren was assigned to 13ql by FISH. Since Ren gene have a VNTR within the 1st intoron of the gene, 5 alleles were detected among 11 inbred strains of rats. Backcross analysis revealed the linkages of Ren with Fh (20cM) and Pep-3 (13cM). The order on chromosome 13 was determined as Fh - Ren - Pep-3. (4) beta2-microglobulin (B2m) : 3 RFLPs by using B2m probe were detected by Southern blot analysis, and backcross analysis revealed the linkage of B2m locus with zi (2 or 9cM), Hao-1 (6cM) , and Srp-1 (29cm) . B2m was assigned to chromosome 3 by a panel of 18 somatic cell hybrid clones made by mouse and rat cells.2. Chromosomal assignment of biochemical genes by somatic cell hybrid clones. We made a panel of 18 somatic cell hybrid clones between mouse and rat cells. This panel enabled us to newly assign 21 biochemical genes of rats on chromosome 1, 2, 3, 4, 7, 8, 9, 10, 12, 13, 14, 19, and 20, respectively. Further Acp-1 and Glo-1 were reconfirmed to locate on chromosome 6 and 20, respectively.

1.DNA探针、回交分析和原位杂交对大鼠基因的染色体定位(1)血管紧张素原(Ang)：通过cDNA和Southern杂交分析，在近交系大鼠中检测到3个Ang等位基因。两组回交分析表明，Ang与其他16个基因座均未连锁，但原位杂交将其定位于19q区。(2)胰岛素-1(Ins-1)：荧光原位杂交(FISH)将INS-1定位于1q55，Southern印迹分析检测到4个等位基因。INS-1与位于1号染色体上的c、Hbb等22个基因座没有连锁，这可能与它们在染色体上的距离较远有关。(3)肾素(REN)：REN被鱼类指定为13ql。由于REN基因在其第一内含子内存在一个VNTR，因此在11个近交系大鼠中检测到5个等位基因。回交分析表明，Ren与Fh(20 CM)和Pep-3(13 CM)有连锁关系。13号染色体上的序列确定为fh-Ren-Pep-3。(4)β2-微球蛋白(B2M)：用B2M探针检测到3个RFLP，回交分析显示B2M与ZI(2 cM或9 cM)、HAO-1(6 CM)和SRP-1(29 CM)连锁。由小鼠和大鼠细胞组成的18个体细胞杂交克隆将B2M定位在3号染色体上。体细胞杂种无性系生化基因的染色体定位。我们在小鼠和大鼠细胞之间制作了一组18个体细胞杂交克隆。这个小组使我们能够分别在1、2、3、4、7、8、9、10、12、13、14、19和20号染色体上新分配21个大鼠的生化基因。进一步证实ACP-1和Glo-1分别位于6号和20号染色体上。