Establishment Of RT-qPCR Method For Detection Of Abortive Transcripts In Vivo And Preliminary Exploration Of Its Biological Functions

Abortion Transcripts(AT)is a non-encoded RNA with only 2 to 10 nt lengths,and AT is generated in the inferior start of the transcription initial(AI)phase.The abortion initiation phenomenon is universal,which occurs in creatures characterized by RNA polymerase,including eukaryotes,prokaryotes,and viruses.In the initial stage of transcription,the RNA polymerase is not quickly and efficiently synthesized,but the RNA constantly synthesized and released in a large amount of 10 nt or less,usually by 10-100 cycles until the RNA above 10 nt can be stabilized Transcription process.The content of the AT in the body is much higher than the full transcriptional m RNA,but because the length of the AT is short,it is caused to detect the current routine RNA quantitative detection method.METHOD: AT is captured using a synthetic capture chain ss DNA(single strand DNA)and two auxiliary chain RNA(B chain and C chain),and then use T4 DNA ligation enzyme to specific repair DNA-RNA hybrid the RNA on the chain is not enhancing but the characteristics of the free RNA cannot be connected,and the heterozygous chain AT is connected to the auxiliary RNA,and it is reversed into c DNA for detection,thereby establishing the RT-q PCR quantitative detection of the AT in the body.method.In order to further explore the function of AT,use the top 30 bits of the initial sequence of apoptotic genes caspase 3 and bax transcriptional start sequences,which use liposome transfection to express these two genes in He La cells.AT,using the established detection technique for Quantitative detection of caspase 3 and bax 8 nt and its maternal gene expression levels of its mother genes(transcriptional genes when transcribed).RESULTS: The muc16 gene 4,6,7,8,10 nt AT homogenous and He La cells were detected by Taq Man-MGB probe method RT-q PCR.muc16 gene 4,6,7,8,10 nt AT that proves that this method can qualitatively and quantitatively detect AT of 4-10 nt in the body.We found that the 8 nt content of 8 nt in the experimental group of transfected plasmid was raised from5.6 and 6.8 times,and bax was up to 12.1 times,although the caspase 3 gene expression did not change,but the AT sequence consistent with it and inhibited the expression of the expression.The raised 1.7 times.It indicates that abortion transcriptional changes in gene expression in transcriptional levels in some way.CONCLUSION: Use this testing technology to quantitatively detect and analyze the abortion transcript of abortion,and provide an important tool for in-depth research on the biological function of abortion transcripts.This study found that the transcription of the inflexible transcriptional transcription of the inflexible induction,indicating that the abundance of abortion may be a new transcriptional regulatory mechanism.