Wilks A F
Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria, Australia.
Proc Natl Acad Sci U S A. 1989 Mar;86(5):1603-7. doi: 10.1073/pnas.86.5.1603.
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.
蛋白质酪氨酸激酶（PTK）在真核细胞生长调节中所起的关键作用，体现在PTK家族成员经常作为生长因子受体或急性转化逆转录病毒的转化因子出现。PTK家族所有成员的一个共同特征是一个高度保守的催化结构域，这是整个家族的特征，并且其活性似乎在细胞内受到PTK其他结构域的严格调控。合成了与所有PTK家族成员催化结构域内两个不变氨基酸序列基序相对应的简并寡核苷酸探针，并用于聚合酶链反应（PCR），以扩增它们之间的cDNA序列。以这种方式从针对小鼠造血细胞系FDC-P1的mRNA制备的cDNA构建了一个M13 PCR文库。然后通过DNA测序筛选该PCR文库以寻找与PTK相关的序列。鉴定出两个序列，根据与已知PTK的序列比较，它们可能编码一类新PTK的代表。
