Peakman T C, Reynolds C H, Willson M G, Moore J D, Spence P, Sydenham M, Linstead D J, Gewert D R, Page M J
Department of Cell Biology, Wellcome Research Laboratories, Beckenham, Kent, U.K.
Biochim Biophys Acta. 1992 Jan 16;1138(1):68-74. doi: 10.1016/0925-4439(92)90153-e.
The cellular gene c-abl is the normal homologue of the transforming gene (v-abl) within the genome of the Abelson leukaemia virus. The cDNA sequence coding for the cellular form of the murine abl gene (c-abl type IV) has been inserted into the baculovirus transfer vector, pAc36C, so that the c-abl gene is under the control of the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus (AcNPV). Spodoptera frugiperda cells infected with the recombinant transfer vector in the presence of wild type AcNPV DNA yielded recombinant, polyhedrin negative virus that expressed moderate levels of the c-Abl protein (representing approx. 0.5-1% of the stained cellular proteins as determined by densitometric scanning). The insect derived c-Abl protein was compared to the P210-BCR/ABL protein from K562 cells, a cell line derived from a patient with chronic myelogenous leukaemia. Antibodies raised against synthetic peptides based on c-abl encoded peptides react with the insect derived c-Abl. In addition, the baculovirus derived c-Abl protein has a tyrosine kinase activity as demonstrated by phosphorylation of a synthetic polypeptide and also by autophosphorylation. Phosphoamino acid analysis of immunoprecipitated, autophosphorylated baculovirus derived c-Abl protein indicates that the majority of label incorporated is on the tyrosine residues. Immunofluorescence microscopy has been used to show that the majority of the c-Abl protein expressed in cells infected with recombinant virus is located in the nuclear and plasma membranes.
细胞基因c-abl是艾贝尔森白血病病毒基因组中转化基因（v-abl）的正常同源物。编码鼠abl基因细胞形式（c-abl IV型）的cDNA序列已被插入杆状病毒转移载体pAc36C中，使得c-abl基因受苜蓿银纹夜蛾核型多角体病毒（AcNPV）多角体蛋白启动子的控制。在野生型AcNPV DNA存在的情况下，用重组转移载体感染草地贪夜蛾细胞，产生了表达中等水平c-Abl蛋白的重组、多角体蛋白阴性病毒（通过密度扫描测定，约占染色细胞蛋白的0.5 - 1%）。将昆虫来源的c-Abl蛋白与来自慢性粒细胞白血病患者的细胞系K562细胞中的P210-BCR/ABL蛋白进行了比较。基于c-abl编码肽段产生的针对合成肽段的抗体与昆虫来源的c-Abl发生反应。此外，杆状病毒来源的c-Abl蛋白具有酪氨酸激酶活性，这通过合成多肽的磷酸化以及自身磷酸化得以证明。对免疫沉淀的、自身磷酸化的杆状病毒来源的c-Abl蛋白进行磷酸氨基酸分析表明，掺入的大部分标记位于酪氨酸残基上。免疫荧光显微镜已用于显示在感染重组病毒的细胞中表达的大多数c-Abl蛋白位于核膜和质膜上。
