Proteomic Analysis of Chloroplast Biogenesis Mutants

Chloroplast mutants in higher plants cause a series of changes in leaf color, while corresponding changes in leaf physiology, encoded genetic characteristics and ultrastructure. Scientists have successfully studied the polypeptide components of chlorophyll proteins of various plants such as snapdragon, tobacco, barley, tomato, and geranium, and elaborated the properties of chloroplast mutant proteins in depth. In addition, several mutant lines defective in chloroplast development have been isolated, and these mutant genes have been found to encode components of several different plastid machinery, including protein import, isoprenoid biosynthesis, RNA processing, protein maturation , plastid gene expression, thylakoid biogenesis, chloroplast to nucleus signal transduction and other important processes. Therefore, the identification of a large number of proteins with chloroplast development and functions is of great significance to study the enzymatic mechanisms and developmental processes of chloroplasts.

Fig. 1. Screening the mutants for chloroplast biogenesis related proteins. (de Luna-Valdez L A, et al., 2014)

Services

Since the expression of genetic information in chloroplasts is often associated with chloroplasts, chloroplast mutants are good materials for studying chloroplast inheritance. Over the years, the team at Lifeasible has extensive experience in proteomic analysis of chloroplast biogenesis mutants. Our scientists have successfully compared the proteomes of four mutants (cla1-1, clb2, clb5, clb19) in Arabidopsis with those of wild-type plants, with the aim of identifying those involved in the regulation of the Arabidopsis chloroplast development of new proteins. In addition, we provide analysis of chlorophyll protein complexes, thylakoid membrane proteins and chloroplast ultrastructure in chloroplast mutant lines of higher plants such as maize and wheat. According to the changes of chloroplast protein separation species, we can test the difference between mutants and normal plants. We mainly adopted the following methods to analyze chloroplast biogenesis mutant proteins:

(1) Isolation and purification of chloroplast proteins.
(2) 2-D PAGE and protein visualization.
(3) In silico processing of gel images.
(4) MALDI-TOF mass spectrometry and protein identification.
(5) In silico analysis of the identified proteins.
(6) Northern blot, gene expression analysis.
(7) Electron microscopy to observe the ultrastructure of chloroplast.

Competitive Advantages

• Potential to study biological processes using comparative proteomic strategies.
• Advanced SDS/PAGE separation and tandem mass spectrometry.
• Team with extensive experience in chloroplast protein research.
• Providing protein analysis of chloroplast mutant lines in a variety of plants.
• Fast and reliable analysis results.

Lifeasible can meet the needs of customers on time and on budget through a wide range of strategies for analyzing chloroplast proteome. Our aim is to be customer-centric and to provide the highest quality service to customers around the world. Our skilled and dedicated scientific researchers ensure that the most appropriate methods and techniques are selected for each specialized chloroplast project. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.

References

1. de Luna-Valdez L A, Martínez-Batallar A G, Hernández-Ortiz M, et al. (2014) Proteomic analysis of chloroplast biogenesis (clb) mutants uncovers novel proteins potentially involved in the development of Arabidopsis thaliana chloroplasts[J]. Journal of proteomics. 111: 148-164.
2. Philippar K, Geis T, Ilkavets I, et al. (2007) Chloroplast biogenesis: the use of mutants to study the etioplast–chloroplast transition[J]. Proceedings of the National Academy of Sciences. 104(2): 678-683.