Hammer J A, Korn E D, Paterson B M
J Biol Chem. 1986 Feb 5;261(4):1949-56.
We have isolated a non-muscle myosin heavy chain gene from Acanthamoeba castellanii using as a heterologous probe a sarcomeric myosin heavy chain gene from Caenorhabditis elegans. The amoeba genomic clone has been tentatively identified as containing a myosin II heavy chain gene based on hybridization to a 5300-nucleotide RNA species, hybrid selection of a mRNA encoding a 185-kDa polypeptide, specific immunoprecipitation of this polypeptide with antiserum to myosin II, and an exact match between the DNA sequence and a carboxyl-terminal myosin II peptide previously sequenced by protein chemical methods (Côté, G.P., Robinson, E.A., Appella, E., and Korn, E. D. (1984) J. Biol. Chem. 259, 12781-12787). We also sequenced a region of the gene whose deduced amino acid sequence shows strong homology with that region of muscle myosins which is thought to be involved in nucleotide binding. These results indicate that the amoeba genomic clone contains at least 90% of the coding information for the 185-kDa heavy chain polypeptide and that the bulk of the gene contains very little intron DNA. Genomic blots of amoeba DNA probed with a portion of this myosin gene indicate the presence of additional highly related sequences within the amoeba genome.
我们使用秀丽隐杆线虫的肌节肌球蛋白重链基因作为异源探针，从卡氏棘阿米巴中分离出了一个非肌肉肌球蛋白重链基因。基于与一种5300个核苷酸的RNA物种的杂交、对编码185 kDa多肽的mRNA的杂交筛选、用肌球蛋白II抗血清对该多肽的特异性免疫沉淀，以及DNA序列与先前通过蛋白质化学方法测序的肌球蛋白II羧基末端肽的精确匹配（Côté, G.P., Robinson, E.A., Appella, E., and Korn, E. D. (1984) J. Biol. Chem. 259, 12781 - 12787），该阿米巴基因组克隆已被初步鉴定为包含肌球蛋白II重链基因。我们还对该基因的一个区域进行了测序，其推导的氨基酸序列与肌肉肌球蛋白中被认为参与核苷酸结合的区域显示出很强的同源性。这些结果表明，该阿米巴基因组克隆包含185 kDa重链多肽至少90%的编码信息，并且该基因的大部分包含很少的内含子DNA。用该肌球蛋白基因的一部分探测阿米巴DNA的基因组印迹表明，阿米巴基因组中存在其他高度相关的序列。
