Arabidopsis transformation and analysis of transgenic plants

Aims

to provide background in the methodologies, aproaches and strategies used for the isolation of a gene(s) to provide the necessary information for the construction of genomic and cDNA libraries to provide information for the use either the genomic or the cDNA library or any other means (pcr methodology) for the isolation of gene(s) to provide the means of analysis for the identification and characterization of the function of an unknown genes by genetics (mutations) or physiological studies to provide experience in the transformation (biotic or abiotic) and the selection and validation of transgenic Arabidopsis by GUS staining and GFP detection

Prerequisites

Molecular Biology
 Genetics
 Biochemistry (less)

Learning Outcomes

The knowledge about the means to isolate genes and the strategies that will be used The characterization of genes by molecular analysis The distinction in molecular versus biochemical approaches The functional characterization of genes and genetics tools students will be able to improve skills on GUS qualitative promoter analysis, acquired knowledge on microscope and GFP detection students will be competent on the scientific notion about the molecular approaches used from the isolation of the gene to the analysis of it  (protein function)

Syllabus

Biochemistry vs. molecular biology, two different biological disciplines with diverged approaches and methodologies used; the genome and the trasncriptome, the genetic information used for the construction of cDNA or genomic libraries; the means to isolate genes from either genomic or cDNA libraries or pcr; the biology of Agrobacterium and biotic transformation; abiotic transformation; vectors used for biotic or abiotic transformation; map based cloning, a genetics approach in isolating genes; verification of transgenes; promoter analysis by GUS; GFP detection in transgenes 

Content Delivery

The method of teaching is power point and practical. First the theoretical background is introduced before the practical course. The logic is to introduce to the students the powerful way to make a transgene based on fundamental scientific approaches and basic science.

Coursework And Assignment Details

There is not enough time (since the unit is given in a very short period of time and students have very limited time) for any assignments in order to evaluate the critical reading or outcome of the students. The learning outcome of the students will be evaluated by written examination at the end of the unit, written assays of about 3-4 questions. These answers will be evaluated and the grades will be given to the students. It will be important for the advancement of the knowledge and critical thinking of the students to give them assignments. However, these assignments should be based not on a single unit but on combination of units (i.e. at the end of, let us say, 4 sequential units)