摘要：碱性亮氨酸拉链（basic Leucine Zipper, bZIP）类转录因子是最古老广泛的真核生物转录因子家族，存在于所有真核生物中。不同bZIP家族成员在功能上既有互补，同时存在差异。在真菌中，bZIP蛋白质参与了调节菌丝生长、多种压力胁迫应答等生理活动，但灵芝中bZIP转录因子的功能尚未被系统地解析。本课题系统地筛选出灵芝bZIP家族转录因子，构建了相应的灵芝bZIP沉默菌株并利用其初步研究了灵芝bZIP转录因子GlbZIP02和GlbZIP08在灵芝生长发育、灵芝三萜合成以及热胁迫、氧化胁迫应答中的作用。结果表明，灵芝bZIP转录因子GlbZIP02和GlbZIP08在灵芝生长发育中发挥必要作用，并对灵芝三萜的合成存在负调控。至于GlbZIP02和GlbZIP08基因是否参与调控灵芝对热胁迫以及氧化胁迫的应答，仍需要进一步研究。25529
毕业论文关键词：碱性亮氨酸拉链；灵芝；灵芝三萜；沉默菌株
The Effects of bZIP Transcription Factors on Growth and Secondary Metabolism in Ganoderma Lucidum
Abstract: The basic leucine zipper (bZIP) proteins are one of the largest and most conserved groups of eukaryotic transcription factors/repressors, found in all eukaryotes. Different members in bZIP family are not only complementary but variant in functions. In fungi, bZIP proteins participate in some physiological activities, such as the regulation of mycelia growth and several kinds of stress response. However, the functions of bZIP transcription factors in Ganoderma lucidum haven’t been analyzed systematically. Here potential bZIP family members of Ganoderma lucidum were screened, and the bZIP family silence strains accordingly were constructed used for preliminary study on the effects of bZIP transcription factors GlbZIP02 and GlbZIP08 on mycelia growth, the biosynthesis of ganoderic acids and the response of heat stress and oxidative stress in Ganoderma lucidum. The results show that the two of bZIP transcription factors GlbZIP02 and GlbZIP08 of Ganoderma lucidum have essential effects on the growth and development of Ganoderma lucidum and play a negative regulation in the biosynthesis of ganoderic acids. As for whether GlbZIP02 and GlbZIP08 regulate the response of heat stress and oxidative stress, further research is necessary.
Key words: basic leucine zipper; Ganoderma lucidum; ganoderic acid; silence strains
目  录
摘要    1
关键词    1
Abstract    1
Key words    1
引言    2
1 材料与方法    2
1.1 实验材料    2
1.1.1 菌株、质粒和试剂    2
1.1.2 菌株培养    2
1.1.3 主要仪器    2
1.2 实验方法    3
1.2.1 灵芝bZIP家族转录因子的系统性筛选     3
1.2.2灵芝bZIP家族转录因子基因的生物信息学分析    3
1.2.3 灵芝bZIP家族转录因子沉默质粒的构建    3
1.2.3.1 灵芝bZIP家族基因沉默片段的PCR扩增    3
1.2.3.2 灵芝bZIP家族基因沉默片段的双酶切    3
1.2.3.3 灵芝bZIP家族基因沉默片段琼脂糖纯化回收    4
1.2.3.4 灵芝bZIP家族基因沉默片段和T载体的连接     4
1.2.3.5 大肠杆菌DH5α感受态的制备和转化    4
1.2.3.6 碱裂解法提取pMD18-T-GlbZIP质粒     5
1.2.3.7 灵芝bZIP家族基因沉默片段和沉默载体的双酶切     5
1.2.3.8 灵芝bZIP家族基因沉默片段和沉默载体酶切片段的琼脂糖纯化回收     5
1.2.3.9 灵芝bZIP家族基因沉默片段和沉默载体酶切片段的连接     5
1.2.3.10 大肠杆菌感受态转化     5 灵芝bZIP转录因子对灵芝生长发育及次生代谢的影响:http://www.751com.cn/shengwu/lunwen_19365.html