Skeletal muscle actin mRNA. Characterization of the 3′ untranslated region

Abstract

Plasmids p749, p106, and p150 contain cDNA inserts complementary to rat skeletal muscle actin mRNA. Nucleotide sequence analysis indicates the following sequence relationships: p749 specifies codons 171 to 360; p150 specifies codons 357 to 374 together with 120 nucleotides of the 3'-non-translated region; p106 specifies the last actin amino acid codon, the termination codon and the entire 3' non-translated region. Plasmid p749 hybridized with RNA extracted from rat skeletal muscle, cardiac muscle, smooth (stomach) muscle, and from brain. It also hybridizes well with RNA extracted from skeletal muscle and brain of dog and chick. Plasmid p106 hybridized specifically with rat striated muscles (skeletal and cardiac muscle) mRNA but not with mRNA from rat stomach and from rat brain. It also hybridized to RNA extracted from skeletal muscle of rabbit and dog but not from chick. Thermal stability of the hybrids and sensitivity to S1 digestion also indicated substantial divergence between the 3' untranslated end of rat and dog skeletal muscle actins. The investigation shows that the coding regions of actin genes are highly conserved, whereas the 3' non-coding regions diverged considerably during evolution. Probes constructed from the 3' non-coding regions of actin mRNAs can be used to identify the various actin mRNA and actin genes.

Differential regulationof glutathione S-transferase Yb1 mRNA levels in rat prostate, liver and brain by androgen

“…As stated in the Materials and Methods, several reference parameters were provided to ensure the quality and quantity of the total RNA and the reproducibility of experiments. Additionally, our results of β-actin gene expression in different tissues agreed with those of other reports [15,16], thus supporting the conviction that there were no problem with RNA loading variations.…”

“…The results are shown in Fig 1. Usually, the mRNA for β-actin was used as an internal control to normalize the tested mRNAs determination. Unfortunately, the steady state of β-actin mRNAs were varied greatly in the tissues tested and even with different size in some tissues as the case of heart, coagulating gland and seminal vesicle which were its cross-hybridizing signals for α-or γ-actin mRNAs [15]. When choosing another popular house-keeping gene GAPDH (Glyceraldehyde-3-phosphate-dehydrogenase) as internal control, the situation seemed to be not improved (Fig 1).…”

cDNA recombinant plasmid complementary to mRNAs for light chains 1 and 3 of mouse skeletal muscle myosin.

“…3, lane c), which gives bands similar to, but fainter than, those seen with mouse muscle (Fig. 3, lane d) (5,(35)(36)(37) and throughout evolution (36,37).…”

Isolation and preliminary characterisation of cDNA clones representing mRNAs associated with tumour progression and metastasis in colorectal cancer

“…RNA dotblot analysis also revealed considerable variation in the abundance of homologous RNAs to the cloned sequences at different stages of tumour progression (Table III). That these differences were not attributable to degradation of the RNA samples concerned was shown by hybridisation of Northern blots to skeletal muscle actin (Shani et al, 1981) Relative abundance ofpLM59-homologous RNA One of the aforementioned recombinants, pLM59, was of particular interest since it appeared to represent a sequence of greater abundance in liver metastases than in primary tumours and normal tissue, and thus may have the potential clinical application as a marker of metastatic disease. The cDNA isolated by EcoRI/BamHI restriction enzyme digest, and labelled by the 'random primed' method of Feinberg and Vogerstein (1983), was used to screen a larger series of RNA dot blots (Table V).…”