[Construction and in vitro activity of specific dual-ribozyme against alpha 1 (I) and (III) procollagen genes].
Overexpression of procollagen gene can cause the extraordinary increase of collagen's synthesis and therefore lead to the keloid and hypertrophic scar. To utilize ribozyme to suppress the expression of procollagen genes, a eukaryotic expression recombinant plasmid containing a dual-ribozyme gene against alpha 1 (I) and alpha 1 (III) procollagen genes was constructed. The ribozyme from in vitro transcription was incubated with target transcripts from recombinant plasmids which separately contained the fragments of the second exons of pro alpha 1 (I) and pro alpha 1 (III) collagen genes under various experimental conditions. The results showed that the dual-ribozyme could efficiently catalyze the specific cleavage of the target RNAs at 37 degrees C, 42 degrees C, 50 degrees C and Mg2+ concentration from 10 mmol/L to 20 mmol/L. This work provided a basis for further study on the ribozyme to suppress the expression of procollagen genes and control the cicatrization.
Kuang
Er-Sheng
ES
College of Life Science, Wuhan University, Wuhan 430072, China.
Yang
Fu-Hua
FH
Wen
Shu-Juan
SJ
Xiang
Kai-Jun
KJ
D001483:Base Sequence; D005091:Exons; D008969:Molecular Sequence Data; D011347:Procollagen / Q000235:genetics*; D012313:RNA / Q000378:metabolism; D016337:RNA, Catalytic / Q000235:genetics* / Q000378:metabolism; D013696:Temperature
D011347:Procollagen; D016337:RNA, Catalytic; D012313:RNA