Zhang Yi, Zhang Xue, Xia Huanzhang, Xue Yuegui, Wang Jianyang, Tian Baozhong, Wei Zhenguo, Lu Changde
School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China.
Acta Biochim Biophys Sin (Shanghai). 2008 Jun;40(6):533-8. doi: 10.1111/j.1745-7270.2008.00425.x.
Cassettes harboring luciferase reporter driven by Bombyx mori cytoplasmic actin gene promoter (A3) (671 bp) and B. mori nuclear polyhedrosis virus immediate-early promoter (IE-1) (580 bp) were transferred to the bacmid AcDeltaEGT to generate the recombinant Autographa californica nuclear polyhedrosis viruses, AcNPVA3Luc and AcNPVIELuc, respectively. Recombinant baculoviruses were injected into the hemocoele of newly ecdysed 5th instar larvae. The activities of the A3 and IE-1 promoters in various tissues were measured by luciferase activity assay and normalized by the copy number of recombinant virus. Results showed that the activity of the A3 promoter was approximately 10-fold higher than the IE-1 promoter. The promoter activities of A3 and IE-1 were highest in the silk gland, followed by fat body, middle gut, malpighian tubule, and hemocyte. In silk gland, activity of the two promoters was highest in posterior silk gland, followed by middle and anterior silk glands. The difference in promoter activities reflects the growth speed of tissue in silkworm larvae. The activity of the A3 promoter remained unchanged and was not inhibited significantly by viral factors at least 3-4 d post injection of rAcNPV.
携带由家蚕细胞质肌动蛋白基因启动子（A3）（671 bp）和家蚕核型多角体病毒立即早期启动子（IE-1）（580 bp）驱动的荧光素酶报告基因的盒式结构被转移到杆粒AcDeltaEGT中，分别产生重组苜蓿银纹夜蛾核型多角体病毒AcNPVA3Luc和AcNPVIELuc。将重组杆状病毒注射到新蜕皮的5龄幼虫的血腔中。通过荧光素酶活性测定法测量A3和IE-1启动子在各种组织中的活性，并通过重组病毒的拷贝数进行标准化。结果表明，A3启动子的活性比IE-1启动子高约10倍。A3和IE-1的启动子活性在丝腺中最高，其次是脂肪体、中肠、马氏管和血细胞。在丝腺中，两个启动子的活性在后丝腺中最高，其次是中丝腺和前丝腺。启动子活性的差异反映了家蚕幼虫组织的生长速度。在注射重组苜蓿银纹夜蛾核型多角体病毒后至少3-4天，A3启动子的活性保持不变，并且未受到病毒因子的显著抑制。
