Allen S L, Zeilinger D, Orias E
Department of Biology, University of Michigan, Ann Arbor 48109-1048, USA.
Genetics. 1996 Dec;144(4):1489-96. doi: 10.1093/genetics/144.4.1489.
We demonstrate a reliable method for mapping conventional loci and obtaining meiotic linkage data for the ciliated protozoan Tetrahymena thermophila. By coupling nullisomic deletion mapping with meiotic linkage mapping, loci known to be located on a particular chromosome or chromosome arm can be tested for recombination. This approach has been used to map three isozyme loci, EstA (Esterase A), EstB (Esterase B), and AcpA (Acid Phosphatase A), with respect to the ChxA locus (cycloheximide resistance) and 11 RAPDs (randomly amplified polymorphic DNAs). To assign isozyme loci to chromosomes, clones of inbred strains C3 or C2 were crossed to inbred strain B nullisomics. EstA, EstB and AcpA were mapped to chromosomes 1R, 3L and 3R, respectively. To test EstA and AcpA for linkage to known RAPD loci on their respective chromosomes, a panel of Round II (genomic exclusion) segregants from a B/C3 heterozygote was used. Using the MAPMAKER program, EstA was assigned to the ChxA linkage group on chromosome 1R, and a detailed map was constructed that includes 10 RAPDs. AcpA (on 3R), while unlinked to all the RAPDs assigned to chromosome 3 by nullisomic mapping, does show linkage to a RAPD not yet assignable to chromosomes by nullisomic mapping.
我们展示了一种可靠的方法,用于绘制传统基因座图谱并获得纤毛原生动物嗜热四膜虫的减数分裂连锁数据。通过将缺体缺失图谱与减数分裂连锁图谱相结合,可以测试已知位于特定染色体或染色体臂上的基因座的重组情况。这种方法已被用于绘制三个同工酶基因座,即酯酶A(EstA)、酯酶B(EstB)和酸性磷酸酶A(AcpA)相对于环己酰亚胺抗性基因座(ChxA)以及11个随机扩增多态性DNA(RAPD)的图谱。为了将同工酶基因座定位到染色体上,将近交系C3或C2的克隆与近交系B的缺体进行杂交。EstA、EstB和AcpA分别被定位到染色体1R、3L和3R上。为了测试EstA和AcpA与它们各自染色体上已知RAPD基因座的连锁关系,使用了来自B/C3杂合子的第二轮(基因组排除)分离群体。使用MAPMAKER程序,EstA被定位到染色体1R上的ChxA连锁群,并构建了一个详细的图谱,其中包括10个RAPD。AcpA(位于3R上)虽然与通过缺体图谱定位到染色体3上的所有RAPD不连锁,但确实与一个尚未通过缺体图谱定位到染色体上的RAPD显示出连锁关系。