Analysis of ABCA Transporter Function in Homologous Recombinant Mice

描述（由申请人提供）：ATP结合盒（ABC）转运蛋白是生物界中最大和最重要的跨膜蛋白家族之一。在ABC家族内，有一组分子被称为A类转运蛋白，它们具有实质性的序列相似性。ABCA类分子包括12种蛋白质，其中只有两种，ABCA 1和ABCA 4，具有明确定义的功能。该授权提出产生缺乏其他10种A类ABCA转运蛋白中的每一种的同源重组小鼠，并表征同源重组小鼠中产生的功能缺陷。一种新的重组细菌人工染色体技术将被用来产生靶向载体和荧光原位杂交将被用来识别正确靶向胚胎干细胞克隆。无效小鼠的表征将采用高通量技术，包括组织脂质的液相色谱质谱分析和基因表达的点样寡核苷酸微阵列测量。在这项工作中产生的所有试剂，包括空小鼠，全长表达cDNA和抗ABCA 1抗体将提供给研究界，以便其他研究人员可以利用它们，以促进我们对这些ABCA转运蛋白在人类生理学和疾病中的作用的理解。

DESCRIPTION (Provided by applicant): The ATP-binding cassette (ABC) transporters compose one of the largest and most important families of transmembrane proteins in the biological kingdom. Within the ABC family is a group of molecules that are referred to as A class transporters that share substantial sequence similarity. The ABCA class molecules comprise twelve proteins of which only two, ABCA1 and ABCA4, have a clearly defined function. This grant proposes to generate homologous recombinant mice lacking each of the other ten A class ABCA transporters and to characterize the resulting functional deficit in the homologous recombinant mice. A novel recombinant bacterial artificial chromosome technology will be used to produce the targeting vectors and fluorescence-in-situ hybridization will be utilized to identify correctly targeted embryonal stem cell clones. Characterization of the null mice will employ high-throughput technologies including liquid chromatography mass spectrometry analysis of tissue lipids and spotted oligonucleotide microarray measurements of gene expression. All of the reagents generated in this work, including null mice, full-length expression cDNAs, and anti-ABCA1 antibodies will be made available to the research community so that other investigators can exploit them in order to facilitate our understanding of the role these ABCA transporters play in human physiology and disease.