PRIMARY STRUCTURE OF AN ALPHA-TUBULIN GENE OF PHYSARUM-POLYCEPHALUM
PRIMARY STRUCTURE OF AN ALPHA-TUBULIN GENE OF PHYSARUM-POLYCEPHALUM
复制标题
DOI：
10.1016/0022-2836(87)90257-9
复制
发表时间：
1987-02-05
影响因子：
5.6
通讯作者：
COX, RA
中科院分区：
文献类型：
--
作者：
MONTEIRO, MJ;COX, RA
一个α-绒泡菌属的微管蛋白基因是作为噬菌体分离的。NM 1149重组体（指定为阶段-NH 4）。N. α.Tu）。Phage-.lambda. N α Tu含有绒泡菌altB基因座的等位基因的4700个碱基对HindIII核DNA片段（四个非连锁α-Tu基因座之一）。微管蛋白基因座）。噬菌体λ的4700碱基对插入片段的亚片段。将N.alpha Tu克隆到噬菌体M13中，并通过双脱氧链终止法确定核苷酸序列。通过引物延伸鉴定转录起始点，并通过S1核酸酶分析定位推定的多聚腺苷酸化位点。将4650个碱基对的HindIII插入噬菌体λ中。N.alpha.Tu跨越完整基因; 5 ''端上游的序列包含RNA转录启动子元件（TATA和CCAAT框）。编码α-半胱氨酸的核苷酸序列微管蛋白含有7个大小为63 - 222个核苷酸的间插序列。外显子具有与绒泡菌α-微管蛋白cDNA克隆，除了三个碱基变化，一个导致瓦尔密码子代替Met密码子，另一个导致Glu密码子代替Asp密码子，第三个变化是沉默的。基因组克隆提供编码cDNA克隆缺失的最后26个氨基酸残基的核苷酸序列。新的序列数据表明α-微管蛋白基因具有C-末端甲硫氨酸密码子，而不是酪氨酸密码子，这在所有α-微管蛋白基因中均被发现。微管蛋白基因测序的日期。
An .alpha.-tubulin gene of Physarum was isolated as a phage-.lambda. NM1149 recombinant (designated phase-.lambda. N.alpha.Tu). Phage-.lambda. N.alpha.Tu contained a 4700 base-pair HindIII nuclear DNA fragment of an allele of the altB locus of Physarum (one of four unlinked .alpha.-tubulin gene loci). Subfragments of the 4700 base-pair insert of phage-.lambda. N.alpha.Tu were cloned into phage M13 and the nucleotide sequence were determined by the dideoxy chain termination method. The start point of transcription was identified by primer extension and a putative polyadenylation site was located by S1 nuclease analysis. The 4650 base-pair HindIII insert into phage-.lambda. N.alpha.Tu spans the complete gene; sequences upstream from the 5'' end contain the RNA transcription promoter elements (the TATA and CCAAT boxes). The nucleotide sequence encoding .alpha.-tubulin contains seven intervening sequences, ranging from 63 to 222 nucleotides in size. The exons have a sequence that is identical with a Physarum .alpha.-tubulin cDNA clone, except for three base changes, one leading to a Val codon in place of a Met codon, another leading to a Glu codon in place of an Asp codon, and the third change is silent. The genomic clone provides the nucleotide sequence coding for the last 26 amino acid residues missing the cDNA clone. The new sequence data indicate that the .alpha.-tubulin gene has a C-terminal methionine codon and not a tyrosine codon, which has been found in all .alpha.-tubulin genes sequenced to date.