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**Introduction**

Protein splicing is a post-translational modification process where inteins, intervening protein sequences, are excised from a precursor protein, and the flanking sequences, exteins, are ligated together. This unique process has been found in all domains of life and plays crucial roles in protein diversity and function. Understanding the mechanisms of protein splicing is essential for uncovering the molecular basis of biological processes and developing novel therapeutic strategies.

**Table of Contents**

1.1 Introduction
1.2 Background of study
1.3 Problem Statement
1.4 Objective of study
1.5 Limitation of study
1.6 Scope of study
1.7 Significance of study
1.8 Structure of the Thesis
1.9 Definition of terms

2. Literature Review
2.1 Historical overview of protein splicing
2.2 Inteins and exteins structures
2.3 Biochemical mechanisms of protein splicing
2.4 Functional significance of protein splicing
2.5 Evolution of protein splicing
2.6 Applications in biotechnology and medicine
2.7 Regulation of protein splicing
2.8 Recent advances in protein splicing research
2.9 Challenges and future directions

3. Research Methodology
3.1 Selection of model systems
3.2 Cloning and expression of precursor proteins
3.3 Site-directed mutagenesis
3.4 Protein purification and characterization
3.5 In vitro splicing assays
3.6 Structural analysis
3.7 Proteomics and mass spectrometry analysis
3.8 Bioinformatics tools for studying protein splicing

4. Discussion of Findings
4.1 Characterization of splicing intermediates
4.2 Identification of key residues for splicing
4.3 Structural insights into splicing mechanisms
4.4 Comparison of different splicing pathways
4.5 Implications for protein engineering
4.6 Crosstalk between splicing and other post-translational modifications
4.7 Regulatory networks of protein splicing
4.8 Future directions and potential applications

5. Conclusion
5.1 Summary of key findings
5.2 Contributions to the field of protein splicing
5.3 Implications for basic research and practical applications
5.4 Future perspectives and directions for further research

**Thesis Overview on Mechanisms of Protein Splicing**

Protein splicing is a remarkable post-translational modification process that involves the removal of inteins from precursor proteins and the subsequent ligation of exteins. This process is catalyzed by the protein splicing machinery, which consists of conserved endonuclease and ligase domains within the inteins. Protein splicing has been found in a wide range of organisms, from bacteria to humans, and is involved in diverse cellular processes, including DNA repair, signal transduction, and immune response.

The aim of this thesis is to provide a comprehensive overview of the mechanisms of protein splicing, including the biochemical and structural aspects, functional significance, evolutionary implications, and potential applications in biotechnology and medicine. The thesis will also discuss the current challenges in studying protein splicing and propose future directions for research in this exciting field.

By conducting a thorough literature review, experimental studies using various model systems, and bioinformatics analyses, this research aims to shed light on the intricate mechanisms underlying protein splicing and its impact on cellular physiology. The findings of this study are expected to enhance our understanding of protein diversity and function, leading to the development of innovative strategies for manipulating protein splicing for therapeutic purposes.

Overall, this thesis will contribute to the growing body of knowledge on protein splicing and provide a solid foundation for further research in this field. The comprehensive analysis of the mechanisms of protein splicing presented in this thesis will lay the groundwork for future studies aimed at unraveling the complexities of this fascinating biological process.

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