Cleveland D W, Hughes S H, Stubblefield E, Kirschner M W, Varmus H E
J Biol Chem. 1981 Mar 25;256(6):3130-4.
Cloned cDNA sequences specific for alpha or beta tubulin mRNAs have been used to show that the multigene families which encode either alpha or beta tubulin are unlinked and dispersed throughout the chicken genome. Fractions of chicken chromosomes partially purified by centrifugation on a sucrose gradient were digested with restriction endonucleases and electrophoresed on agarose gels. The DNA was transferred to nitrocellulose filters and hybridized to labeled probes constructed from cloned cDNA sequences specific for alpha or beta tubulin. We find alpha tubulin sequences on four different chicken chromosomes and beta tubulin sequences on at least two different chromosomes. Moreover, using chicken chromosomes further purified with a fluorescent cell sorter, we have been able unambiguously to localize alpha tubulin genes to chromosome 1 and chromosome 8 and two of the beta genes to chromosome 2.
已使用针对α或β微管蛋白mRNA的克隆cDNA序列来表明,编码α或β微管蛋白的多基因家族是不连锁的,且分散在整个鸡基因组中。通过在蔗糖梯度上离心部分纯化的鸡染色体片段,用限制性内切酶消化,并在琼脂糖凝胶上进行电泳。将DNA转移到硝酸纤维素滤膜上,并与由针对α或β微管蛋白的克隆cDNA序列构建的标记探针杂交。我们在四条不同的鸡染色体上发现了α微管蛋白序列,在至少两条不同的染色体上发现了β微管蛋白序列。此外,使用通过荧光细胞分选仪进一步纯化的鸡染色体,我们能够明确地将α微管蛋白基因定位到1号染色体和8号染色体,将两个β基因定位到2号染色体。