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2. 发明申请

WO1993022331A1 HUMAN CRABP-I AND CRABP-II 审中-公开
标题翻译：人CRABP-I和CRABP-II
公开(公告)号：WO1993022331A1
公开(公告)日：1993-11-11
申请号：PCT/US1993003936
申请日：1993-04-27
申请人： THE REGENTS OF THE UNIVERSITY OF MICHIGAN , ASTROM, Anders , VOORHEES, John, J. , PETTERSON, Ulrika , TAVAKKOL, Amir
发明人： THE REGENTS OF THE UNIVERSITY OF MICHIGAN
IPC分类号： C07H21/04
CPC分类号： C12Q1/6897 , C07K14/70567 , G01N2333/70567 , G01N2500/00
摘要： The sequences encoding two isoforms of human cellular retinoic acid binding proteins, CRABP-I and CRABP-II, have been cloned and sequenced and are set forth with their corresponding amino acid sequences in SEQ ID NOS. 1-4. The indentification of human CRABP nucleic and amino acid sequences provides the basis for the construction of recombinant human CRABP vectors and expression constructs. Human CRABP can also be synthesized or produced ex vivo, e.g. in bacterial or other production systems. Ligand binding assays, including recombinant and chimeric receptor reporter assays, and direct and competition hybridization assays employing the human CRABP sequences herein described can be used to test drugs for retinoic induction and tissue specificity for pathologies in which retinoids are implicated. Immunoassays utilizing antibodies or binding fragments produced to human CRABP can also be used to test patient tissues for the presence and levels of CRABP for diagnosis and to monitor treatment. The indentification of the nucleic and amino acids sequences for human CRABP-I and CRABP-II also contributes to the elucidation of the function and interaction of the retinoid-binding proteins. The CRABP-II gene, isolated from a human placenta genomic library, spans 6 kilobases and includes 4 exons. One major transcription initiation site was mapped to an A residue 137 nucleotides upstream of the ATG initiation codon. CRABP-II mRNA was rapidly induced within 2-6 hours in culture by retinoic acid, primarily due to an increased rate of transcription which required on-going synthesis. The human CRABP-II gene is thus apparently transcriptionally regulated by a newly synthesized regulator protein. Once the CRABP-II is produced, message stabilization may provide means by which elevated CRABP-II in mRNA is maintained.
摘要翻译：编码人类细胞视黄酸结合蛋白（CRABP-1和CRABP-II）的两种同工型的序列已被克隆并测序，并以SEQ ID NOS中相应的氨基酸序列进行阐述。 1-4。人类CRABP核酸和氨基酸序列的鉴定提供了构建重组人类CRABP载体和表达构建体的基础。人类CRABP也可以离体合成或产生，例如，在细菌或其他生产系统中。配体结合测定法，包括重组和嵌合受体报告基因测定，以及使用本文所述的人类CRABP序列的直接和竞争杂交测定法，可用于检测视黄酸诱导药物和组胺特异性，其中涉及类维生素A。利用针对人类CRABP产生的抗体或结合片段的免疫测定法也可用于测试患者组织中CRABP的存在和水平以进行诊断和监测治疗。人CRABP-1和CRABP-II的核酸和氨基酸序列的鉴定也有助于阐明类视黄醇结合蛋白的功能和相互作用。从人胎盘基因组文库分离的CRABP-II基因跨越6千碱基并且包括4个外显子。一个主要的转录起始位点被映射到ATG起始密码子上游137个核苷酸的A残基。 CRABP-II mRNA在视黄酸培养2-6小时内迅速诱导，主要是由于需要持续合成的转录速率增加。因此，人类CRABP-II基因显然被新合成的调节蛋白转录调节。一旦CRABP-II产生，信号稳定可以提供维持mRNA中升高的CRABP-II的方式。

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