Kropyvko S. Alternative splicing and alternative transcription in formation of human іntersectіn 1 gene transcripts diversity

The object - regulation of gene expression. The aim - to define the role of alternative splicing and alternative transcription in creation of isoform diversity of human multifunction adapter protein ITSN1. Methods - methods to molecular biology methods which include a polymerase chain reaction following the reverse transcription (RT-PCR) for the analysis of expression, cloning of full-length DNA fragments, 5'RACE-analysis, Southern-blot analysis, methods of cell biology, in particular, cultivation of human cell lines, luciferase test, bioinformatics analysis of DNA sequences. Results and novelty - dissertation is based on the analysis of the diversity of alternative spliced forms of human intersectin 1 (ІTSN1) transcripts, search and characterization of alternative promoters in ІTSN1 gene structure. Three new alternative splicing events of the long form of human ІTSN1 have been identified and expression analysis of these forms in human tissues has been performed. Alternative splicing events in 5'UTR of ІTSN1 mRNA and its expression have been analyzed. Fifteen ІTSN1 transcripts with full-length coding sequence generated as the result of alternative splicing have been cloned. Using bioinformatic methods the main promoter and 5'UTR of ІTSN1 mRNA were described. Alternative promoter of human ІTSN1 gene located at intron 5 have been identified and characterized. Expression analysis of the transcripts synthesized from this promoter has been performed. Using deletion mutants critical promoter region of alternative promoter has been identified and transcripts with full-length coding sequence generated from this promoter have been cloned and analyzed.