AI Article Synopsis
- A 1 kb scaffold-attachment region (SAR) from the silkworm’s rRNA gene was cloned into a vector for studying gene expression.
- The SAR increased luciferase gene expression by up to 15 times in stable cell lines but had no significant effect in transient transfections.
- The SAR likely requires integration into chromosomal DNA and binds to nuclear matrix proteins, which may be crucial for its role in enhancing transcription.
Article Abstract
The 1 kb scaffold-attachment region (SAR) at 5' non-transcription region of rRNA gene of silkworm Attacus ricini was cloned into eukaryotic expression vector pLu, which contained luciferase report gene and neo(R) selecting marker. After transfection of constructs into NIH3T3 cell line by using cation liposome, the luciferase activity was monitored to check the SAR's function. The results demonstrated that the SAR could enhance gene expression up to 15-fold in stable transformed cells, but no obvious gene expression was observed in transient transfection. Its effect on gene expression appeared to requirechromosomal integration. Southwestern blotting experiments showed that SAR specifically bound to nuclear matrix proteins of NIH3T3 cells. The binding with nuclear matrix may be necessary for SAR function of transcriptional enhancement.
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