An analysis of the presence of proteins on different levels in dark grown and light grown plants : Biogenesis of organells

Project Report from the year 2014 in the subject Biology - Botany, grade: 1,6, LMU Munich, language: English, abstract: In the following experiments the aim is to analyse the presence of proteins on different levels, comparing dark grown and light grown plants. In part A the samples are being analysed on transcript level, determining the amount of transcripts for LHC, POR and ALB4. The author also compares transcription levels of those proteins in Wild-type plants and dsbA mutants, which have PSII deficiency. For controlling the 18S rRNA is being used, which is a housekeeping gene and should be equally expressed in all samples. In part B there will be a comparison between the same samples on protein level by protein isolation, Coomassie staining and immunostaining with antibodies. Here the author also focuses on the proteins ALB4, LHC and POR. Furthermore, in organelle labelling was performed, which should show differences in protein levels between chloroplasts and etioplasts. Part C is focusing on the light-harvesting complexes, which developed during de-etoilation of dark grown seedlings. A semi-native gel electrophoresis was performed and chlorophyll a and chlorophyll b amounts were measured. As samples the wild-type, dsbA and alb3 mutants were used and compared to each other. In these experiments the aim is to analyse the presence of those proteins on different levels, comparing dark grown and light grown plants. In part A the samples are being analysed on transcript level, determining the amount of transcripts for LHC, POR and ALB4. Here we also compare transcription levels of those proteins in Wild-type plants and dsbA mutants, which have PSII deficiency. As a control the 18S rRNA is being used, which is a housekeeping gene and should be equally expressed in all samples. In part B we will compare the same samples on protein level by protein isolation, Coomassie staining and immunostaining with antibodies. Here we also focus on the proteins ALB4, LHC and POR. Furthermore, in organelle labelling was performed, which should show differences in protein levels between chloroplasts and etioplasts. Part C is focusing on the light-harvesting complexes, which are developed during de-etoilation of dark grown seedlings. A semi-native gel electrophoresis was performed and chlorophyll a and chlorophyll b amounts were measured. As samples the wild-type, dsbA and alb3 mutants were used and compared to each other.