Guénette S, Prichard R K, Klein R D, Matlashewski G
Institute of Parasitology, McGill University, Macdonald College, Ste-Anne de Bellevue, Quebec, Canada.
Mol Biochem Parasitol. 1991 Feb;44(2):153-64. doi: 10.1016/0166-6851(91)90001-m.
A genomic clone containing a beta-tubulin gene from the parasitic nematode Brugia pahangi was isolated. This gene was sequenced to determine its size, structural organization, and corresponding primary amino acid sequence. The coding sequence of the beta-tubulin gene spans 3.8 kb, is organized into 9 exons and expresses an mNRA of 1.8 kb which codes for a protein of 448 amino acids. The predicted beta-tubulin amino acid sequence is 89%, 94%, 90% and 88% identical to the chicken beta 2, and the Caenorhabditis elegans ben-1, tub-1 and mec-7 gene products, respectively. Southern hybridization analyses demonstrated that there is only one copy of this gene isotype but that other distinct beta-tubulin genes may exist in the Brugia pahangi genome. A nematode specific antipeptide rabbit antiserum raised against the predicted amino acid sequence of the extreme carboxy-terminal region of the B. pahangi beta-tubulin was used to identify beta-tubulin isoforms in adult nematodes and microfilariae. Isoforms detected by this nematode-specific antipeptide antiserum were identical in both adult worms and microfilariae and did not differ from the isoform patterns detected by a monoclonal antibody recognizing a conserved beta-tubulin epitope. This suggests that this carboxy-terminal peptide is highly represented in the beta-tubulin isoforms of B. pahangi.
从寄生线虫彭亨布鲁线虫中分离出一个含有β-微管蛋白基因的基因组克隆。对该基因进行测序以确定其大小、结构组织和相应的初级氨基酸序列。β-微管蛋白基因的编码序列跨度为3.8kb，由9个外显子组成，表达一个1.8kb的mRNA，编码一个448个氨基酸的蛋白质。预测的β-微管蛋白氨基酸序列与鸡的β2、秀丽隐杆线虫的ben-1、tub-1和mec-7基因产物分别有89%、94%、90%和88%的同一性。Southern杂交分析表明，该基因同型仅有一个拷贝，但彭亨布鲁线虫基因组中可能存在其他不同的β-微管蛋白基因。针对彭亨布鲁线虫β-微管蛋白极端羧基末端区域的预测氨基酸序列产生的一种线虫特异性抗肽兔抗血清，用于鉴定成虫线虫和微丝蚴中的β-微管蛋白同工型。这种线虫特异性抗肽抗血清检测到的同工型在成虫和微丝蚴中是相同的，并且与识别保守β-微管蛋白表位的单克隆抗体检测到的同工型模式没有差异。这表明该羧基末端肽在彭亨布鲁线虫的β-微管蛋白同工型中高度表达。
