A new 5′ terminal murine GAPDH exon identified using 5′RACE LaNe

  title={A new 5′ terminal murine GAPDH exon identified using 5′RACE LaNe},
  author={Daniel J. Park},
  journal={Molecular Biotechnology},
In this work, a ligation-independent, fully gene-specific, nested polymerase chain reaction (PCR) method for the elucidation of 5′ cDNA sequence is described and demonstrated for the first time. Two manifestations of the method, rapid amplification of cDNA ends (RACE) by lariat-dependent nested PCR 5′ (RACE LaNe), at least as simple to perform as conventional RACE, were successfully applied to the murine housekeeping genes phosphoglycerate kinase 1 (PGK1), β-actin (β-ACT), and glyceraldehyde-3… Expand
6 Citations
Lariat-dependent nested PCR for flanking sequence determination.
Methods detailed in this chapter relate to the use of Lariat-dependent Nested (LaNe) PCR to characterize unknown RNA or DNA sequence flanking known regions. A multitude of approaches designed toExpand
The polymerase chain reaction (PCR) is a scientific technique in molecular biology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands toExpand
Complete genomic sequence of an infectious pancreatic necrosis virus isolated from rainbow trout (Oncorhynchus mykiss) in China
The sequence of ChRtm213 was the first gene sequence of IPNV isolates in China, and the results suggest that the Chinese IPNV isolate has relative closer relationship with Japanese IPNV strains. Expand
Characterisation of the marsupial-specific ATRY gene: implications for the evolution of male-specific function.
The isolation and sequencing of the tammar wallaby ATRY cDNA, and comparison of its sequence with that of tammar ATRX show the conservation and loss of particular motifs identify those required for testicular function (ATRY) and function in other tissues (ATRX). Expand


3' RACE LaNe: a simple and rapid fully nested PCR method to determine 3'-terminal cDNA sequence.
3′ RACE LaNe is demonstrated: a simple and rapid fully nested PCR method to determine 3′-terminal cDNA sequence that outperforms a conventional heminested anchored PCR 3′ Race series when it is applied to the long and complex tammar wallaby ATRX gene and the mouse PGK1 and GAPD genes. Expand
CapSelect: a highly sensitive method for 5' CAP-dependent enrichment of full-length cDNA in PCR-mediated analysis of mRNAs.
The method combines the 5'-CAP-dependent addition of specifically three to four non-templated dCMP residues to the 3'-end of full-length cDNAs by reverse transcriptases in the presence of manganese and the controlled ribonucleotide tailing of cDNA ends by terminal deoxynucleotidyl transferase using rATP. Expand
Ligation-mediated amplification of RNA from murine erythroid cells reveals a novel class of beta globin mRNA with an extended 5'- untranslated region
The evolutionary conservation of this relationship suggests the importance of the GATA promoter element of the mouse beta globin gene and its possible involvement in developmental regulation of expression of this gene. Expand
Amplification and characterization of a β-globin gene synthesized in vitro
It was possible to show that the synthetic globin DNA is a faithful copy of β-globin mRNA by comparing the nucleotide sequences known or predicted from the amino acid sequences of α- and β-chain rabbit hemoglobin. Expand
Enzymatic in vitro synthesis of globin genes
Reassociation, hybridization, and restriction endonuclease studies, as well as electrophoretic analyses, indicate that the sequential actions of reverse transcriptase, DNA polymerase 1, and S1 nuclease generate full-length, double-stranded synthetic globin genes. Expand
A general method for cloning eukaryotic structural gene sequences.
The susceptibility of inserted beta globin, sequences to the restriction endonuclease EcoRI confirms the existence of a site already found through previous nucleotide sequence analysis, as well as estimating the amount of inserted globin sequences. Expand
Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer.
  • M. Frohman, M. Dush, G. Martin
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1988
The efficacy of this cDNA cloning strategy was demonstrated by isolating cDNA clones of mRNA from int-2, a mouse gene that expresses four different transcripts at low abundance, the longest of which is approximately 2.9 kilobases. Expand
Stepwise biosynthesis in vitro of globin genes from globin mRNA by DNA polymerase of avian myeloblastosis virus.
  • F. Rougeon, B. Mach
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1976
Data indicate that a loop structure on the 3' end of cDNA allowed DNA synthesis to take place by a "self-priming" mechanism, and some of the double-stranded DNA synthesized corresponded to the entire sequence of the 9S mRNA template. Expand
New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase.
  • M. Sirover
  • Biology, Medicine
  • Biochimica et biophysica acta
  • 1999
The mechanisms through which mammalian cells may utilize GAPDH amino acid sequences to provide new functions and to determine its intracellular localization are considered and the interrelationship between new GAPDh activities and its role in cell pathologies is addressed. Expand
High-efficiency full-length cDNA cloning by biotinylated CAP trapper.
We have devised a method for efficiently constructing high-content full-length cDNA libraries based on chemical introduction of a biotin group into the diol residue of the cap structure of eukaryoticExpand