Two putative protein-tyrosine kinases identified by application of the polymerase chain reaction.
Two putative protein-tyrosine kinases identified by application of the polymerase chain reaction.
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通过应用聚合酶链式反应鉴定出两种推定的蛋白酪氨酸激酶。
DOI：
10.1073/pnas.86.5.1603
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发表时间：
1989
影响因子：
11.1
通讯作者：
A. Wilks
中科院分区：
文献类型：
--
作者：
A. Wilks
蛋白酪氨酸激酶（PTK）在真核细胞的生长调节中发挥的关键作用表现在PTK家族成员作为生长因子受体或作为急性转化逆转录病毒的转化剂的频繁出现。PTK家族所有成员的共同特征是高度保守的催化结构域，其是整个组的特征，并且其活性似乎在细胞内受到PTK的其他结构域的严格调节。简并的寡核苷酸探针，对应于两个不变的氨基酸序列基序内的催化结构域的所有PTK家族成员的合成和聚合酶链反应（PCR）扩增它们之间的cDNA序列。以这种方式从针对来自鼠造血细胞系FDC-P1的mRNA制备的cDNA产生M13 PCR文库。然后通过DNA测序筛选PCR文库中的PTK相关序列。基于与已知PTK的序列比较，鉴定了两个序列，其可以编码一类新的PTK的代表。
The pivotal role that protein-tyrosine kinases (PTKs) play in the growth regulation of eukaryotic cells is manifest in the frequent appearance of members of the PTK family as growth factor receptors or as the transforming agents of acutely transforming retroviruses. A feature common to all members of the PTK family is a highly conserved catalytic domain which is characteristic of the group as a whole and whose activity appears to be tightly regulated within the cell by other domains of the PTK. Degenerate oligonucleotide probes corresponding to two invariant amino acid sequence motifs within the catalytic domains of all PTK family members were synthesized and employed in the polymerase chain reaction (PCR) to amplify cDNA sequences between them. An M13 PCR library was produced in this way from cDNA prepared against mRNA from the murine hemopoietic cell line FDC-P1. The PCR library was then screened by DNA sequencing for PTK-related sequences. Two sequences were identified that, on the basis of sequence comparison with known PTKs, may encode representatives of a new class of PTK.