Splice junctions, determination

The term splicing refers to the process by which introns are removed and the mRNA put back together to form a continuous coding sequence in the 5 -3 direction. Remembering how accurate this process must be is important. If only a single nucleotide of an intron were left in the processed mRNA, the protein made from that mRNA would be non-functional, because the ribosome would read the wrong codons. The cellular machinery that splices pre-mRNAs uses information at the splice junctions to determine where to cut and where to rejoin the mRNA. Removal of introns from transcripts containing more than one intron usually occurs in a preferred but not exclusive order. Several pathways are used. 

The structure of the cutinase gene has been determined using genomic DNA cloned in X-phage. This gene contains one 51 bp intron which has the typical junction and splicing signals... 

In addition, we would like to mention that the oligomer 2. was designed to be complementary to the splice acceptor junction of the tat HIV gene vide infra) in order to evaluate its inhibitory capacity on HIV-infected MT4 cells. As such experiments are done in culture medium (S days at 37°C in RPMI 1640 containing 10% of heat-inactivated calf serum), we have been able to determine its stability under such stringent conditions using the ISRP on-line HPLC approach that we recently introduced. 

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