The successful cloning of recombinants between cDNA from fractionated poly(A)+-RNA of Brockmann bodies of the carp and the plasmid pBR322 in Escherichia coli chi 1776 is reported. One of the recombinant clones has been identified as a preproinsulin-cDNA recombinant by the hybrid-arrest translation assay. Recombination was at the PstI site of pBR322; reconstitution of this site was by 3'-tailing of the vector with dGn. The transformants were screened by in situ hybridization with kinase-labeled poly(A)+-RNA sedimenting at 9S from Brockmann bodies. Restriction analysis was performed on 26 of the strongly hybridizing clones to estimate the size of the inserted cDNA. Six of the recombinants studied contain inserts of a size approximating to full length 9S preproinsulin mRNA. The hybrid-arrest translation assay on selected clones identified one as a recombinant containing the preproinsulin cDNA sequence.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/0378-1119(90)90325-l | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
PAX4 preserves endoplasmic reticulum integrity preventing beta cell degeneration in a mouse model of type 1 diabetes mellitus.
Diabetologia
April 2016
Pancreatic Islet Development and Regeneration Unit, Department of Stem Cells, Andalusian Center for Molecular Biology and Regenerative Medicine (CABIMER), Avda Américo Vespucio, Parque Científico y Tecnológico Cartuja 93, 41092, Seville, Spain.
Aims/hypothesis: A strategy to enhance pancreatic islet functional beta cell mass (BCM) while restraining inflammation, through the manipulation of molecular and cellular targets, would provide a means to counteract the deteriorating glycaemic control associated with diabetes mellitus. The aims of the current study were to investigate the therapeutic potential of such a target, the islet-enriched and diabetes-linked transcription factor paired box 4 (PAX4), to restrain experimental autoimmune diabetes (EAD) in the RIP-B7.1 mouse model background and to characterise putative cellular mechanisms associated with preserved BCM.
View Article and Find Full Text PDFEffect of hepatic insulin expression on lipid metabolism in diabetic mice.
J Diabetes
May 2016
Division of Pediatric Endocrinology, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
Background: Hypertriglyceridemia is a common lipid disorder that is characterized by elevated plasma levels of triglyceride (TG)-rich particles, such as very low-density lipoprotein (VLDL), in poorly controlled diabetes. The aim of the present study was to determine the potential therapeutic effect of hepatic insulin production on hypertriglyceridemia in mice.
Methods: Mice were induced diabetic and hypertriglyceridemic by streptozotocin (STZ) treatment.
Full-length cDNA cloning and structural characterization of preproinsulin in Alligator sinensis.
Genet Mol Res
October 2014
College of Life Sciences, Anhui Normal University, Wuhu, Anhui, China.
Insulin is an important endocrine hormone that plays a critical physiological role in regulating metabolism and glucostasis in vertebrates. In this study, the complete cDNA of Alligator sinensis preproinsulin gene was cloned for the first time by reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends methods; the amino acid sequence encoded and protein structure were analyzed. The full-length of preproinsulin cDNA sequence consists of 528 base pairs (bp), comprising a 34-bp 5'-untranslated region, a 170-bp 3'-untranslated region and an open reading frame that is 324 bp in length.
View Article and Find Full Text PDFAutoimmunity against INS-IGF2 protein expressed in human pancreatic islets.
J Biol Chem
October 2013
From the Department of Clinical Sciences, Lund University Diabetes Center, Lund University, Skåne University Hospital SUS, SE-205 02 Malmö, Sweden and.
Insulin is a major autoantigen in islet autoimmunity and progression to type 1 diabetes. It has been suggested that the insulin B-chain may be critical to insulin autoimmunity in type 1 diabetes. INS-IGF2 consists of the preproinsulin signal peptide, the insulin B-chain, and eight amino acids of the C-peptide in addition to 138 amino acids from the IGF2 gene.
View Article and Find Full Text PDFConstruction of Plasmid Insulin Gene Vector Containing Metallothionein IIA (pcDNAMTChIns) and Carbohydrate Response Element (ChoRE), and Its Expression in NIH3T3 Cell Line.
Int J Endocrinol Metab
July 2013
Department of Biochemistry and Nutrition, School of Medicine, Hamadan University of Medical Science, Hamadan, IR Iran.
Background: Type 1 diabetes mellitus is one of the metabolic diseases that cause insulin-producing pancreatic ß cells be destroyed by immune system self-reactive T cells. Recent-ly, new treatment methods have been developed including use of the stem cells, ß islet cells transplantation and gene therapy by viral and non-viral gene constructs.
Objectives: The aim of this project was preparing the non-viral vector containing the glucose inducible insulin gene and using it in the NIH3T3 cell line.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!