PubMed:25982022 JSONTXT

{"project":"silkworm","denotations":[{"id":"T1","span":{"begin":41,"end":47},"obj":"Gene:100127099"},{"id":"T2","span":{"begin":112,"end":121},"obj":"Species:7091"},{"id":"T3","span":{"begin":177,"end":183},"obj":"Gene:100127099"},{"id":"T4","span":{"begin":268,"end":276},"obj":"Species:7091"},{"id":"T5","span":{"begin":352,"end":358},"obj":"Gene:100127099"},{"id":"T6","span":{"begin":647,"end":656},"obj":"Species:7091"},{"id":"T7","span":{"begin":832,"end":841},"obj":"Species:7091"},{"id":"T8","span":{"begin":954,"end":963},"obj":"Species:7091"},{"id":"T9","span":{"begin":990,"end":999},"obj":"Species:7091"},{"id":"T10","span":{"begin":1112,"end":1118},"obj":"Gene:100127099"},{"id":"T11","span":{"begin":1280,"end":1288},"obj":"Species:7091"},{"id":"T12","span":{"begin":1339,"end":1348},"obj":"Species:7091"},{"id":"T13","span":{"begin":1433,"end":1439},"obj":"Gene:100127099"},{"id":"T14","span":{"begin":1486,"end":1492},"obj":"Gene:100127099"}],"text":"The 5'-UTR intron of the midgut-specific BmAPN4 gene affects the level and location of expression in transgenic silkworms.\nIntrons are important for regulating gene expression. BmAPN4, which has a 5'-UTR upstream intron (5 UI), is specifically expressed in the entire silkworm midgut. In our previous study, the promoter region upstream of the 5 UI of BmAPN4 was cloned and identified as the P3 promoter (P3P) with activity only in the anterior midgut. In this study, the sequence consisting of the P3P and the 5 UI was cloned and named as P3P+5 UI. A transgenic vector was constructed in which EGFP was controlled by P3P+5 UI. Transgenic P3+5 UI silkworms were generated by embryo microinjection. RT-PCR showed P3P+5 UI activity throughout the larval stage. Intense green fluorescence was seen only in the entire midgut of P3+5 UI silkworms and expression was confirmed by RT-PCR. qPCR revealed that expression of EGFP in the anterior midgut of P3+5 UI silkworms was 64% higher than in P3 silkworms, indicating the 5 UI sustained intron-mediated enhancement of gene expression. These results suggested that the BmAPN4 5 UI affected the level and site of expression. The 5 UI was cloned and added behind P2P, another specific promoter with activity only in the anterior midgut of silkworm, to construct the P2P+5 UI and transgenic P2+5 UI silkworms. Expression patterns were the same for P2P+5 UI and P2P, suggesting that the 5UI of BmAPN4 did not affect P2P. This study found that the BmAPN4 5 UI affected the amount and location of gene expression. Its influence appeared to be dependent on a specific promoter."}

{"project":"silkwormbase","denotations":[{"id":"T1","span":{"begin":41,"end":47},"obj":"Gene:100127099"},{"id":"T10","span":{"begin":1112,"end":1118},"obj":"Gene:100127099"},{"id":"T11","span":{"begin":1280,"end":1288},"obj":"Species:7091"},{"id":"T12","span":{"begin":1339,"end":1348},"obj":"Species:7091"},{"id":"T13","span":{"begin":1433,"end":1439},"obj":"Gene:100127099"},{"id":"T14","span":{"begin":1486,"end":1492},"obj":"Gene:100127099"},{"id":"T2","span":{"begin":112,"end":121},"obj":"Species:7091"},{"id":"T3","span":{"begin":177,"end":183},"obj":"Gene:100127099"},{"id":"T4","span":{"begin":268,"end":276},"obj":"Species:7091"},{"id":"T5","span":{"begin":352,"end":358},"obj":"Gene:100127099"},{"id":"T6","span":{"begin":647,"end":656},"obj":"Species:7091"},{"id":"T7","span":{"begin":832,"end":841},"obj":"Species:7091"},{"id":"T8","span":{"begin":954,"end":963},"obj":"Species:7091"},{"id":"T9","span":{"begin":990,"end":999},"obj":"Species:7091"}],"text":"The 5'-UTR intron of the midgut-specific BmAPN4 gene affects the level and location of expression in transgenic silkworms.\nIntrons are important for regulating gene expression. BmAPN4, which has a 5'-UTR upstream intron (5 UI), is specifically expressed in the entire silkworm midgut. In our previous study, the promoter region upstream of the 5 UI of BmAPN4 was cloned and identified as the P3 promoter (P3P) with activity only in the anterior midgut. In this study, the sequence consisting of the P3P and the 5 UI was cloned and named as P3P+5 UI. A transgenic vector was constructed in which EGFP was controlled by P3P+5 UI. Transgenic P3+5 UI silkworms were generated by embryo microinjection. RT-PCR showed P3P+5 UI activity throughout the larval stage. Intense green fluorescence was seen only in the entire midgut of P3+5 UI silkworms and expression was confirmed by RT-PCR. qPCR revealed that expression of EGFP in the anterior midgut of P3+5 UI silkworms was 64% higher than in P3 silkworms, indicating the 5 UI sustained intron-mediated enhancement of gene expression. These results suggested that the BmAPN4 5 UI affected the level and site of expression. The 5 UI was cloned and added behind P2P, another specific promoter with activity only in the anterior midgut of silkworm, to construct the P2P+5 UI and transgenic P2+5 UI silkworms. Expression patterns were the same for P2P+5 UI and P2P, suggesting that the 5UI of BmAPN4 did not affect P2P. This study found that the BmAPN4 5 UI affected the amount and location of gene expression. Its influence appeared to be dependent on a specific promoter."}