拟南芥VPS34回补酵母Vps34突变体的构建.doc

文档介绍：
拟南芥 VPS34 回补酵母 Vps34 突变体的
构建#
章永春,龚清秋**
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(南开大学生命科学学院,天津 300071)
摘要:磷脂酰肌醇 3-激酶在动物与酵母细胞中参与调控自噬与液泡蛋白分选等胞内运输过
程,然而其在植物中的作用还不明确。我们在前期研究中发现拟南芥的磷脂酰肌醇 3-激酶
(AtVps34)可能参与营养回收,然而此作用是否通过自噬来完成尚有待研究。我们通过同
源重组在可以通过测定碱性磷酸酶 ALP 活性而定量测定自噬的酵母菌株中敲除 ScVps34 基
因,然后将 AtVps34 克隆到 pRS414 载体上,转入酵母,获得了 AtVps34 回补酵母 vps34 突
变体的回补株系。该材料为后续研究 AtVps34 在自噬中的可能作用提供了基础。
关键词:植物学;Vps34;自噬;酵母回补
中图分类号:[Q291]
Complementation of the yeast vps34 mutant with AtVPS34
ZHANG Yongchun, GONG Qingqiu
(College of Life Sciences, Nankai University, TianJin 300071)
Abstract: Phosphotidylinositol 3-kinases (PtdIns 3-kinases) are involved in many intracellular
trafficking processes in mammalian and yeast, including autophagy and vacuolar protein sorting.
The function of PI3K in plant is still unclear. We have previously found that AtVps34 is
associated with nutrient recycling. Whether it is achieved through autophagy remains elusive.
Here, a plementation stain was generated. Homologous bination was used to
knock out ScVps34 in a yeast strain TN124, a strain for quantifying autophagy through alkaline
phosphatase activity assay. Then AtVps34 was cloned into vector pRS414 for transformation of
knockout yeast strains. The strains will serve as materials for future research on AtVps34.
Keywords: Plant biology; Vps34; autophagy; plementation
0 引言
细胞自噬(Autophagy)是目前已知最高效的亚细胞降解途径[1]。在这一途