Font Size: a A A

Cloning And Function Analysis Of Annexin5 In Arabidopsis Thaliana

Posted on:2012-01-14Degree:MasterType:Thesis
Country:ChinaCandidate:G WeiFull Text:PDF
GTID:2120330335470673Subject:Botany
Abstract/Summary:PDF Full Text Request
Annexin are Ca2+ and phospholipid binding proteins forming an evolutionary conserved multigene family, with members of the family being expressed throughout animal and plant kingdoms. It is generally assumed that Annexins are implicated in several processes related to membranes, including regulation of membrane organisation, membrane trafficking, secretion and vesicular trafficking.But we are still far from a comprehensive description of Annexin function in cell processes. There is hardly any research or report about the Annexin5 of higher plant. Therefore, this research project is to use molecular biology and cell biology methods, the first attempt to study Arabidopsis pollen-specific expression of the membrane protein family members-Annexin5. Results are as follows:1. In order to investigate the gene function of Annexin5, the full length of cDNA was cloned and inserted into the plant expression vector of pCAMBIA1300,then fused to GFP gene. We also constructed a set of plant expression vector include mutational or deleted position.for example, mutational in Ca2+ binging position,mutational in PIP2 binging position. In this study,We use pollen special promoter LAT52 and ZM13.2. The recombination vector was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens through the floral dip method. Filtered by hygromyein, meanwhile, fluoroscopic examination proved that the plant expression vectors had been integrated into arabidopsis genome. Now we have got all the transgenic plants.3. Screening transgenic plants stably expressed and observed its phenotype, now we have obtained homozygotes of 1300-lat52-GFP and 1300-lat52-GFP-Ann5. We confirmed that whether the overexpression of Annexin5 has influences on the pollen germination,pollen tube growth and actin cytoskeleton. By observe the effect of Annexin5 on Arabidopsis thaliana pollen tube growth, we wanted to reveal the molecular mechanism of membrane protein involved in pollen germination and pollen tube growth.This study laid a foundation for revealed the molecular mechanism of members of the annexin family, Which act as Hub protein play an important role in the interactions among membrane, actin cytoskeleton and calcium ions during polar growth of pollen tube. has important theoretical value and application prospect.
Keywords/Search Tags:Annnexin5, mutant, pollen tube Growth, actin cytoskeleton
PDF Full Text Request
Related items