Biomaterial Database

[The labeling of procollagen gene probe by PCR and the detection of procollagen gene expression in hepatic stellate cells]. 2001

W X Zhao, and C L Liang, and B Zhao, and R Q Pang, and Z L Chen

Medical Laboratory of Kunming General Hospital, Chengdu Command, Kunming 650032, China.

OBJECTIVE To establish a new method for labeling of procollagen gene probe and the detection of procollagen mRNA expression in HSC. METHODS According to gene sequences from NCBI Gene Bank, the primers for the amplification of type I, III and IV procollagen genes were designed by OLIGO software, the procollagen genes were amplified by RT-PCR and were labeled by PCR and DIG-dUTP, and the probes were applied to detect procollagen mRNA expression in cultured hepatic stellate cells by in situ hybridization. RESULTS The procollagen genes were successfully amplified by the Primers and RT-PCR and were labeled by DIG-dUTP and PCR, the procollagen mRNA expression in cultured hepatic stellate cells was detected by the probes. CONCLUSIONS A new simplified method for labeling of procollagen genes is successfully found and it can be used for other gene amplification and labeling.

UI	MeSH Term	Description	Entries
D011347	Procollagen	A biosynthetic precursor of collagen containing additional amino acid sequences at the amino-terminal and carboxyl-terminal ends of the polypeptide chains.	Protocollagen,Procollagen Type M
D002478	Cells, Cultured	Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.	Cultured Cells,Cell, Cultured,Cultured Cell
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D015342	DNA Probes	Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections.	Chromosomal Probes,DNA Hybridization Probe,DNA Probe,Gene Probes, DNA,Conserved Gene Probes,DNA Hybridization Probes,Whole Chromosomal Probes,Whole Genomic DNA Probes,Chromosomal Probes, Whole,DNA Gene Probes,Gene Probes, Conserved,Hybridization Probe, DNA,Hybridization Probes, DNA,Probe, DNA,Probe, DNA Hybridization,Probes, Chromosomal,Probes, Conserved Gene,Probes, DNA,Probes, DNA Gene,Probes, DNA Hybridization,Probes, Whole Chromosomal
D015870	Gene Expression	The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.	Expression, Gene,Expressions, Gene,Gene Expressions
D016133	Polymerase Chain Reaction	In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.	Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain
D055166	Hepatic Stellate Cells	Perisinusoidal cells of the liver, located in the space of Disse between HEPATOCYTES and sinusoidal endothelial cells.	Ito Cells,Cell, Hepatic Stellate,Cells, Hepatic Stellate,Cells, Ito,Hepatic Stellate Cell,Stellate Cell, Hepatic,Stellate Cells, Hepatic