基于Codon优化的全基因扫描识别了Bombyx mori核多面体病毒多面体过度表达所必需的隐藏核酸
Susumu Katsuma1, Noriko Matsuda-Imai1
1Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Journal of molecular biology
|May 9, 2024
概括
在Bombyx mori核聚合体病毒 (BmNPV) 中,对多面体 (POLH) 基因的共优化减少了POLH积累. 在5'编码区域的特定核酸序列对于高POLH表达和闭塞体形成至关重要.
科学领域:
- 分子生物学分子生物学
- 病毒学 病毒学
- 昆虫病理学 昆虫病理学
背景情况:
- 阿尔法巴库洛病毒通过多面体 (POLH) 过度表达产生闭塞体 (OBs).
- 波尔基因的促进物对baculovirus表达系统至关重要.
- 在POLH积累中,polh编码序列的作用尚不清楚.
研究的目的:
- 调查子使用和核酸序列对POLH积累的影响.
- 为了在多个基因中识别特定的序列元素,对过度表达至关重要.
主要方法:
- 在BmNPV.中产生了代码子优化的polh突变 (co-polh).
- 分析了受感染细胞中的POLH积累和POLHmRNA水平.
- 构建了嵌合体的波兰突变体,并进行了 luciferase 记者测试.
主要成果:
- 由于对polh的优化,POLH的积累显著降低.
- 减少的POLH水平归因于减少的polh mRNA,而不是翻译问题.
- 一个30bp 5'近端编码区域对于高的mRNA水平至关重要.
- 这一区域的五个特定的核酸差异被确定为过度表达的关键.
结论:
- 在polh编码区域内的核酸序列,特别是5'区域,对于过度表达至关重要.
- 单独优化Codon并不能保证增加蛋白质的积累.
- 确定了有效的alphabaculovirus基因表达的关键cis作用元素.
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