| :Rodents have played important role in maintaining the structure and function of biocoenose and the flow of material and energy among ecosystem. However, in recent years, rodents have turned to be more and more dangerous for most habitats.it has caused more and more attention for its high reproductive capacity.4.5SRNA is arodent-specific small noncoding nuclearRNA. As an atrotransposon, 4.5SRNA exist in the rat genomes of repeat sequences in short, scattered in individual development process with dynamic change and aging process closely related。So the study of the copies of 4.5 SRNA appears especially important important。SYBR Green I FQ-PCR characterized by quick,specificity, sensitivity provides a profitable method for detecing 4.5S RNA and quantitating.Total RNA was extracted from the liver tissue of Cricetulus barabensis, the cDNA of 4.5SRNA was amplified by reverse transcription-polymerase chain reaction method, then constructed with the vector PMD-18 and sequenced. It will be restructured plasmid purification as standard quality grain.standard quality grain was tested by the aging double dilution methods, the transgene copy numbers in transgenic mice were determined by quantitative real-time PCR.The conserved mouse housekeeping gene, GAPDH, served as an internal control to calculate transgene copy numbe.The method was highly specific and sensitive,and could be used for rapid quantitative detection of the copies of 4.5SRNA. The copies of Rattusnorvegicus, Musmusculus, Cricetulusbarabensis,Apodemusagrarius, Cricetulus triton respectively was 2570,1096,1513,1770,954 through Real time fluorescence quantitative polymerase chain reaction (PCR) method。The experiment confirms that 4.5SRNA exatly exits in thegenome of animal, Further illustrates the biological significance of research reversal seat.the reason that the atrotransposon 4.5 SRNA only exit in in the anmial genomes,how it resoles,and the function in thelife metabolism are still to be further discussed. |
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