Akella R, Arasu P, Vaidya A B
Department of Microbiology and Immunology, Hahnemann University, Philadelphia, PA 19102-1192.
Mol Biochem Parasitol. 1988 Aug;30(2):165-74. doi: 10.1016/0166-6851(88)90109-0.
By using a Trypanosoma brucei alpha-tubulin cDNA probe under reduced stringency hybridization conditions, we have isolated two genomic clones that constitute portions of alpha-tubulin genes of the rodent malarial parasite Plasmodium yoelii. P. yoelii has two alpha-tubulin genes, the 3' portions of which were present in the two clones, Py alpha T1 and Py alpha T2, containing 1.3 kb and 6.6 kb EcoRI fragments respectively. The 1358 bp Py alpha T1 clone was completely sequenced and found to contain 591 nucleotides of uninterrupted coding sequence with a strong bias for AT-rich codons, starting with codon 254 of a consensus alpha-tubulin sequence. Numerous attempts to clone 5' portions of these genes were unsuccessful. A single mRNA of 2.3 kb was recognized by both the clones in the erythrocytic stages of P. yoelii. A probe constituting the untranslated sequences of Py alpha T1 also recognized this RNA but failed to hybridize with Py alpha T2 sequences, indicating that the gene represented by the Py alpha T1 clone was expressed during the erythrocytic stages. The deduced amino acid sequence of the Py alpha T1 gene terminates in Tyr-Glu instead of Glu-Tyr observed in alpha-tubulins of almost all other organisms. The difference observed may have implications for alpha-tubulin metabolism in malarial parasites.
在较低严谨性杂交条件下,通过使用布氏锥虫α-微管蛋白cDNA探针,我们分离出了两个基因组克隆,它们构成了啮齿动物疟原虫约氏疟原虫α-微管蛋白基因的部分。约氏疟原虫有两个α-微管蛋白基因,其3'部分存在于两个克隆PyαT1和PyαT2中,分别包含1.3 kb和6.6 kb的EcoRI片段。对1358 bp的PyαT1克隆进行了全序列测定,发现其包含591个不间断的编码序列核苷酸,对富含AT的密码子有强烈偏好,从共有α-微管蛋白序列的第254个密码子开始。多次尝试克隆这些基因的5'部分均未成功。在约氏疟原虫的红细胞阶段,两个克隆都识别出一条2.3 kb的单一mRNA。构成PyαT1非翻译序列的探针也识别了这种RNA,但未能与PyαT2序列杂交,这表明PyαT1克隆所代表的基因在红细胞阶段表达。PyαT1基因推导的氨基酸序列以Tyr-Glu结尾,而不是几乎所有其他生物的α-微管蛋白中观察到的Glu-Tyr。观察到的这种差异可能对疟原虫中的α-微管蛋白代谢有影响。