Analysis Of MiRNA-1 And MiRNA-133 Expression In Muscle Tissues From Different Porcine Species

MicroRNAs (miRNAs) are a class of small molecule of non-coding RNA about 20-25 bp, which are generally offered by Dicer enzyme from a hairpin precursor secondary structure by RNA processing, can trigger the degradation of target mRNA and / or the translation inhibition to mediate post-transcriptional gene silencing. RNAs of these small molecules often expressed by the method of preventing recurrent or developmental stage-specific expression of specific, they play an important role in regulating the organization, cell growth,development,differentiation,apoptosis and function. They exist in a wide range of eukaryotes, many studies have proved that these microRNA has a very important regulatory function in vivo, almost all the function of the level of cells. People focused their attention on the study of traditional and protein synthesis of the three RNA (mRNA, tRNA and rRNA) in the past, and hundreds of miRNA were identified from the nematodes and human bodies since the small RNA (mainly include two types of miRNA and siRNA) were found;Plants of the Arabidopsis genome analysis showed that, miRNA role most associated with the development of the transcription factor, in particular with the establishment of phenotype and cell differentiation.MiRNA have been extensively studied in animals and plants as an important regulatory factors in vivo of eukaryotic recently. With the development of experimental and bioinformatics methods, more and more miRNA was cloned, but the number of miRNA be identified is far from the saturation value of their theory in all species, now majority of miRNA identified are highly conserved evolutionarily by the experiments in mammals. Recent studies have shown that a large number of non-conserved miRNA are existed in higher organisms. Therefore, they are of great significance in the continuation of the new experimental miRNA identification, expression and functional studies, so it is still a hot research direction to the isolation and identification of miRNA expression and function of miRNA.MiRNA-1 and miRNA-133 was first cloned and identified from the muscle tissue of mice and human, with a length of 22nt, from the more than 70 nt precursor stem-loop structure shear processing maturity, and their coding genes located in the 2 and 18 chromosomes, cluster distribution, the common transcription. Study shows that miRNA-1 and miRNA-133 present the temporal and spatial expression of a certain specificity in human, mouse, such as skeletal muscle cells during differentiation and maturation. In skeletal muscle, miRNA-1 can promote myogenesis by muscle targets HDAC4, and miRNA-133 increased myoblast rate of cell proliferation through the inhibition of serum response factor (serum response factor, SRF), suggesting that both can play an important role in the regulation of proliferation and differentiation of skeletal muscle.This paper studies that explore the application of real-time quantitative PCR, quantitative measurement of miRNA-1 and miRNA-133 in pig skeletal muscle in the feasibility of the expression level and explore the different varieties of pig skeletal muscle the expression the different species of adult pigs as a model. Subjects divided into two parts.: Part I: Construction the plasmid vector of the miRNA-1 and miRNA-133 and real-time fluorescence quantitative PCR to establishment detection methods of the quantitative of a mature miRNA. The study proves that miRNA-1 and miRNA-133 exist the expression in pig skeletal muscle, determine the consistency of the mature sequence between human and mouse, and this also proves that the miRNA have the highly conserved; partâ…¡: This study detects the expression of miRNA-1 and miRNA-133 by RT-PCR between the same and different pigs(Blue Pool pigs, Landrace, White Flower), and analysis of the results of expression. The results showed that their expression has a significant difference between miRNA-1 and miRNA-133 in their skeletal muscle. The quality of the meat may have a certain degree of correlation between miRNA-1 and miRNA-133 from experimental data and test results of pork quality indicators.In this study detects the expression of miRNA-1 and miRNA-133 in skeletal muscle tissue by real-time quantitative PCR, and analysis the significance of development of pig skeletal muscle in light of the actual data. The study provide new ideas and clues to research miRNA regulatory mechanism in muscle growth, the formation of quality traits, to dig new target gene of genetic engineering breedings in muscle production, quality and to improve molecular of meat production traits.