Evolution of the functional human beta-actin gene and its multi-pseudogene family: conservation of noncoding regions and chromosomal dispersion of pseudogenes.

Abstract

We have assigned six members of the human beta-actin multigene family to specific human chromosomes. The functional gene, ACTB, is located on human chromosome 7, and the other assigned beta-actin-related sequences are dispersed over at least four different chromosomes including one locus assigned to the X chromosome. Using intervening sequence probes, we showed that the functional gene is single copy and that all of the other beta-actin related sequences are recently generated in evolution and are probably processed pseudogenes. The entire nucleotide sequence of the functional gene has been determined and is identical to cDNA clones in the coding and 5' untranslated regions. We have previously reported that the 3' untranslated region is well conserved between humans and rats (Ponte et al., Nucleic Acids Res. 12:1687-1696, 1984). Now we report that four additional noncoding regions are evolutionarily conserved, including segments of the 5' flanking region, 5' untranslated region, and, surprisingly, intervening sequences I and III. These conserved sequences, especially those found in the introns, suggest a role for internal sequences in the regulation of beta-actin gene expression.

Antisense ‘knockdowns’ of M1 receptors induces transient anterograde amnesia in mice

“…RNase treated before RT or reverse transcriptase-omitted samples were the negative PCR controls in order to exclude any possible contamination by endogenous genomic DNA and/or by amplified DNA carry-over. Primers 5%-GCG GGA AAT CGT GCG TGA CAT-3%; residues 2104-2125 and 5%-GAT GGA GTT GAA GGT AGT TTC GTG-3%; residues 2409-2432, according to the published sequence (Ng et al, 1985), were used in the quantitative protocol for the amplification of i-actin cDNA as an internal standard (Quattrone et al, 1995b). For both primer pairs, the three-step PCR cycles consisted of 1 min denaturation at 92°C, 1 min annealing at 58°C and 1 min extension at 72°C.…”

Reversible antisense inhibition of Shaker-like Kv1.1 potassium channel expression impairs associative memory in mouse and rat

“…Forward and reverse primers were upstream from and downstream to the segment of Kv1.1 mRNA targeted by the antisense ODNs. To control for the integrity of RNA and for differences attributable to errors in experimental manipulation from tube to tube, primers for rat phosphoglycerate kinase 1 (PGK1) (19) or mouse ␤-actin (20) were included in the PCR reactions and generated a 183-and 232-bp PCR product, respectively. Oligonucleotide sequences and location of the primers were as follows: 5Ј-AGGTGCT-CAACAACATGGAG-3Ј (PGK1, residues 777-796), 5Ј-TACCAGAGGCCACAGTAGCT-3Ј (PGK1, residues 940-959), 5Ј-GCGGGAAATCGTGCGTGACATT-3Ј (␤-actin, residues 2106-2127) and 5-GATGGAGTTGAAGGTAGTT-TCGTG-3Ј (␤-actin, residues 2409-2432).…”

Genomic organization, splice variants and expression of CGM1, a CD66‐related member of the carcinoembryonic antigen gene family

“…The intactness of the RNA samples was confirmed by analyzing for the presence of p-actin mRNA. This reaction was carried out both with and without reverse transcription prior to PCR to detect any genomic DNA contamination with processed j?-actin pseudogenes (Ng et al, 1985). The j?-actin cDNA fragments, which were obtained after reverse transcription, are shown (Fig.…”