A cloned chromogranin A (CgA) cDNA detects a 2.3Kb mRNA in diverse neuroendocrine tissues |
| |
| Authors: | L J Deftos S S Murray D W Burton R J Parmer D T O'Connor A M Delegeane P L Mellon |
| |
| Institution: | 1. University of California, San Diego, California, USA;2. San Diego VA Medical Center, San Diego, California, USA;1. Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan;2. Department of Chemistry, Faculty of Physical Sciences, Imo State University, PMB2000 Owerri, Nigeria;3. Department of Chemical Engineering, Federal University of Petroleum Resources, PMB 1221 Effurun, Nigeria;1. Department of Physiology, Cardiovascular Research Center, Center for Muscle Biology, University of Kentucky, Lexington, KY, USA;2. Cellular and Molecular Arrhythmia Research Program, University of Wisconsin, Madison, WI, USA;3. Department of Cardiology, Gill Heart Institute, University of Kentucky, Lexington, KY, USA;1. Department of Engineering, University of Cambridge, Cambridge CB3 0FA, United Kingdom;2. Wireless and Photonic Networks Research Centre, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia;3. Department of Electronic Engineering, Faculty of Engineering and Green Technology, Universiti Tunku Abdul Rahman, 31900 Kampar, Perak, Malaysia;4. Department of Physics, Faculty of Science, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia;5. Department of Electrical Engineering, College of Engineering, P.O. Box 800, King Saud University, Riyadh 11421, Saudi Arabia;1. Cerealicultura, Facultad de Ciencias Agrarias y Forestales, Universidad Nacional de La Plata, Av. 60 y 119, La Plata, Argentina;2. CONICET CCT La Plata, Calle 8 N° 1467, La Plata, Buenos Aires, Argentina;3. Departamento de Producción Vegetal, Cátedra de Cerealicultura, Universidad de Buenos Aires, Av. San Martín 4453, C1417DSE Buenos Aires, Argentina;4. IFEVA-CONICET, Universidad de Buenos Aires, Av. San Martín 4453, C1417DSE Buenos Aires, Argentina;1. BG Klinikum Unfallkrankenhaus Berlin gGmbH, Achenbach-Krankenhaus Königs Wusterhausen, Alice Salomon Hochschule Berlin, Deutschland;2. BG Klinikum Unfallkrankenhaus Berlin gGmbH, Hochschule der Deutschen Gesetzlichen Unfallversicherung (HGU), Deutschland;3. Deutsche Gesetzliche Unfallversicherung (DGUV) Spitzenverband der gewerblichen Berufsgenossenschaften und der Unfallversicherungsträger der öffentlichen Hand, Deutschland;4. BG Klinikum Unfallkrankenhaus Berlin gGmbH, Universitätsmedizin Greifswald, Deutschland |
| |
| Abstract: | CgA is a 72Kd protein of unknown function that is present in many neuroendocrine tissues and co-secreted with their resident hormones. We prepared a cDNA library to the mRNA from CgA-producing human medullary thyroid carcinoma (MTC) cells in the expression vector lambda gt11. The library was screened with a panel of one polyclonal and two monoclonal antibodies to CgA. The specificity of the antibodies for CgA was demonstrated by immunoassay, immunohistology, and immunoprecipitation of the in vitro translation products of mRNA from CgA-producing tissues. A chromogenic second antibody identified five immunoreactive clones. Their cDNA inserts were isolated after EcoRI digestion and agarose gel electrophoresis. These cDNAs were 32P-labelled and used as probes in Northern hybridization studies. An mRNA of approximately 2.3Kb was detected with the cDNA probes in human cell lines from MTC and lung cancers that were shown to produce CgA and in human pheochromocytoma and bovine adrenal medulla tissue. To confirm its identity, one of the putative CgA cDNAs was subcloned into a plasmid and was used to hybridization-arrest the in vitro translation of CgA mRNA. These studies demonstrate the cloning of cDNAs which hybridize with CgA mRNA from diverse neuroendocrine tissues. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
