Narang S K, Seawright J A
ARS, Insects Affecting Man and Animals Research Laboratory, Gainesville, FL 32604.
J Am Mosq Control Assoc. 1993 Jun;9(2):147-9.
The histone genes of Anopheles albimanus were mapped by in situ hybridization to 6 bands in Region 34A on the right arm of chromosome 3. A genomic library was made by cloning fragments of 15 to 23 kb (derived from partial EcoRI digestion) into the phage vector, EMBL4, and probed with the histone gene repeat of Drosophila melanogaster. Thirty-two phages containing histone gene sequences were isolated from about 10(5) plaque-forming units (pfu). Complete EcoRI digestion of DNA from 5 of the 32 recombinant phages and the genomic DNA of An. albimanus yielded a single 3.84-kb fragment that contained sequences homologous to the 5 histone genes of D. melanogaster. This 3.84-kb unit of mosquito histone genes was subcloned into puc19 plasmid, and the resulting clone (palbi34A) was used for in situ hybridization to salivary gland chromosomes.
通过原位杂交技术,将白线按蚊的组蛋白基因定位到了3号染色体右臂34A区域的6条带纹上。构建了一个基因组文库,方法是将15至23 kb的片段(源自部分EcoRI酶切)克隆到噬菌体载体EMBL4中,并用黑腹果蝇的组蛋白基因重复序列进行探针杂交。从大约10⁵个噬菌斑形成单位(pfu)中分离出了32个含有组蛋白基因序列的噬菌体。对32个重组噬菌体中的5个以及白线按蚊的基因组DNA进行完全EcoRI酶切,产生了一个单一的3.84 kb片段,该片段包含与黑腹果蝇5个组蛋白基因同源的序列。这个3.84 kb的蚊虫组蛋白基因单位被亚克隆到puc19质粒中,所得克隆(palbi34A)用于对唾液腺染色体进行原位杂交。