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Key points 4-5 Changing the natural gene sequences

Previously we have learned that gene mutations can be generated by various factors, while our body is smart enough to have various damage repair systems to fight for them. In this lecture, we will learn the site-directed mutagenesis that can change the natural sequence of DNA controlably. Exploring the relationship of changes in gene and amino acid sequences with the structure and function of mutant protein, will provide an effective method to elucidate the micro-level mechanism of gene regulation and disease pathogenesis. So what are the main technologies for site-directed mutagenesis? How can they achieve precise mutagenesis? Let's start our new journay with these questions!

 

Key points:

1. The principle and applications of site-directed mutagenesis

2. Methods of site-directed mutagenesis

Ø  Oligonucleotide primer-mediated site-directed mutagenesis

Ø  PCR mediated site-directed mutagenesis

Ø  Cassette mutagenesis

3. The Principle and protocols for oligonucleotide primer-mediated site-directed mutagenesis                  

4. Applications of homologous recombination in gene editing

Ø  Gene knockout

Ø  Gene knock-in

下一节:Video 4-5 Changing the natural gene sequences

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Basic Molecular Biology 课程列表：

Chapter 1 The material basis of genetic information

- C1-Lecture 1 The past and present journey of molecular biology

--Key points C1-L1 The past and present journey of molecular biology

--Video-C1-L1 The past and present journey of molecular biology

--Discussion C1-L1

--Lecture 1-Homework

-C1-Lecture 2 The nature of genetic material

--Key points C1-L2 The nature of genetic material

--Video-C1-L2 The nature of genetic material

--Discussion C1-L2

--C1-Lecture 2-Homework

-C1-Lecture 3 Amazing DNA double helix

--Key points C1-L3 Amazing DNA double helix

--Video C1-L3 Amazing DNA double helix

--Discussion C1-L3

--C1-Lecture 3-Homework

-C1-Lecture 4 Supercoiled DNA

--Key points C1-L4 Supercoiled DNA

--Video C1-L4 Supercoiled DNA

--Discussion C1-L4

--C1-Lecture 4-Homework

-C1-Lecture 5 Denaturation, renaturation and hybridization

--Key points C1-L5 Denaturation, renaturation and hybridization

--Video C1-L5 Denaturation, renaturation and hybridization

--Discussion C1-L5

--C1-Lecture 5-Homework

-C1-Lecture 6 Nucleic acids extraction and gel electrophoresis

--Key points C1-L6 Nucleic acid extraction and gel electrophoresis

--Video C1-L6 Nucleic acid extraction and gel electrophoresis

--Discussion C1-L6

--C1-Lecture 6-Homework

-Chapter 1 Test

-Learning resources

--Web resources

-- References

Chapter 2 From gene to chromosome

-C2-Lecture 1 Genome does not equal to chromosome

--Key points C2-L1 Genomes are not equal to chromosomes

--Video C2-L1 Genomes are not equal to chromosomes

--Discussion C2-L1 Genomes are not equal to chromosomes

--C2-Lecture 1-Homework

-C2-Lecture 2 Nucleosomes and their Assembly

--Key points C2-L2 Nucleosomes and their Assembly

--Video C2-L2 Nucleosomes and their Assembly

--Discussion C2-L2 Nucleosomes and their Assembly

--C2-Lecture 2-Homework

-C2-Lecture 3 Higher order chromatin structure

--Key points C2-L3 Higher order chromatin structure

--Video C2-L3 Higher order chromatin structure

--Discussion C2-L3 Higher order chromatin structure

--C2-Lecture 3-Homework

-C2-Lecture 4 Chromatin modification and remodeling

--Key points C2-L4 Chromatin modification and remodeling

--Video C2-L4 Chromatin modification and remodeling

--Discussion C2-L4 Chromatin modification and remodeling

--C2-Lecture 4-Homework

-Chapter 2 Tests

-Learning resources

--Web resources

--References

Chapter 3 Who is in pairs? DNA replication

-C3-Lecture 1 Semiconservative DNA replication

--Key points C3-L1 Semiconservative DNA replication

--Video C3-L1 Semiconservative DNA replication

--Discussion C3-L1 Semiconservative DNA replication

--C3-Lecture1 Homework

-C3-Lecture 2 The mechanism of DNA polymerase

--Key points C3-L2 The mechanism of DNA polymerase

--Video C3-L2 The mechanism of DNA polymerase

--Discussion C3-L2 The mechanism of DNA polymerase

--C3-Lecture 2-Homework

-C3-Lecture 3 Process of DNA replication

--Key points C3-L3 Process of DNA replication

--Video C3-L3 Process of DNA replication

--Discussion C3-L3 Process of DNA replication

--C3-Lecture 3-Homework

-C3-Lecture 4 Regulation of DNA replication initiation

--Key points C3-L4 Regulation of DNA replication initiation

--Video C3-L4 Regulation of DNA replication initiation

--Discussion C3-L4 Regulation of DNA replication initiation

--C3-Lecture 4-Homework

-C3-Lecture 5 The end replication problem and telomere

--Key points C3-L5 The end replication problem and telomere

--Video C3-L5 The end replication problem and telomere

--Discussion C3-L5 The end replication problem and telomere

--C3-Lecture 5-Homework

-C3-Lecture 6 Let’s “cook” DNA

--Video C3-L6 Let’s “cook” DNA

--Discussion C3-L6 Let’s “cook” DNA

--C3-Lecture 6-Homework

- Chapter 3 Tests

-Learning resources

--Web resources

--References

Chapter 4 DNA mutation and repair

-C4-Lecture 1 Replication errors and the mismatch repair system

--Key points 4-1 Replication errors and the mismatch repair system

--Video 4-1 Replication errors and the mismatch repair system

--Discussion 4-1 Replication errors and the mismatch repair system

--C4-Lecture 1-Homework

-C4-Lecture 2 DNA damage

--Key points 4-2 DNA damage

--Video 4-2 DNA damage

--Discussion 4-2 DNA damage

--C4-Lecture 2-Homework

-C4-Lecture 3 DNA Repair

--Key points 4-3 DNA Repair

--Video 4-3 DNA Repair

--Discussion 4-3 DNA Repair

--C4-Lecture 3-Homework

-C4-Lecture 4 DNA Homologous Recombination

--Key points 4-4 DNA Homologous Recombination

--Video 4-4 DNA Homologous Recombination

--Discussion 4-4 DNA Homologous Recombination

--C4-Lecture 4-Homework

-C4-Lecture 5 Changing the natural gene sequences

--Key points 4-5 Changing the natural gene sequences

--Video 4-5 Changing the natural gene sequences

--Discussion 4-5 Changing the natural gene sequences

-Chapter 4 test

- Learning resources

--Web resources

--References

--References

Chapter 5 Transcription in prokaryotes

-C5-Lecture 1 RNA polymerase

--Key points 5-1 RNA polymerase

--Video 5-1 RNA polymerase

--Discussion 5-1 RNA polymerase

--C5-Lecture 1-Homework

-C5-Lecture 2 Transcription process in prokaryotes

--Key points 5-2 Transcription process in prokaryotes

--Video 5-2 Transcription process in prokaryotes

--Discussion 5-2 Transcription process in prokaryotes

--C5-Lecture 2-Homework

-C5-Lecture 3 Transcriptional regulation in bacterial I: Lac operon

--Key points 5-3 Transcriptional regulation in bacterial I: Lac operon

--Video 5-3 Transcriptional regulation in bacterial I: Lac operon

--Discussion 5-3 Transcriptional regulation in bacterial I: Lac operon

--C5-Lecture 3-Homework

-C5-Lecture 4 Transcriptional regulation in bacterial II: Tryptophan operon

--Key points 5-4 Transcriptional regulation in bacterial II: Tryptophan operon

--Video 5-4 Transcriptional regulation in bacterial II: Tryptophan operon

--Discussion 5-4 Transcriptional regulation in bacterial II: Tryptophan operon

--C5-Lecture 4-Homework

-Chapter 5 Test

-Learning resources

--References

Chapter 6 Transcription in eukaryotes

-Lecture 1 Eukaryotic RNA polymerase and promoter

--Key points 6-1 Eukaryotic RNA polymerase and promoter

--Video 6-1 Eukaryotic RNA polymerase and promoter

--Discussion 6-1 Eukaryotic RNA polymerase and promoter

--C6-Lecture 1-Homework

-Lecture 2 Transcription process in eukaryotic cell

--Key points 6-2 Transcription process in eukaryotic cell

--Video 6-2 Transcription process in eukaryotic cell

--Discussion 6-2 Transcription process in eukaryotic cell

--C6-Lecture 2-Homework

-Lecture 3 Transcription factors of eukaryotic cell

--Key points 6-3 Transcription factors of eukaryotic cell

--Video 6-3 Transcription factors of eukaryotic cell

--Discussion 6-3 Transcription factors of eukaryotic cell

--C6-Lecture 3-Homework

-Lecture 4 Epigenetic Regulation of gene expression

--Key points 6-4 Epigenetic Regulation of gene expression

--Video 6-4 Epigenetic Regulation of gene expression

--Discussion 6-4 Epigenetic Regulation of gene expression

--C6-Lecture 4-Homework

-Chapter 6 Test

-Learning resources

--Web resources

--References

Chapter 7 RNA Processing

-Lecture 1 RNA splicing

--Key points 7-1 RNA splicing

--Video 7-1 RNA splicing

--Discussion 7-1 RNA splicing

--C7-Lecutre 1-Homework

-Lecture 2 Cotranscriptional processing of nuclear pre-mRNA

--Key points 7-2 Cotranscriptional processing of nuclear pre-mRNA

--Video 7-2 Cotranscriptional processing of nuclear pre-mRNA

--Discussion 7-2 Cotranscriptional processing of nuclear pre-mRNA

--C7-Lecture 2-Homework

-Lecture 3 One gene one protein? Alternative RNA splicing

--Key points 7-3 One gene one protein? Alternative RNA splicing

--Video 7-3 One gene one protein? Alternative RNA splicing

--Discussion 7-3 One gene one protein? Alternative RNA splicing

--C7-Lecture 3-Homework

-Lecture 4 “Seeing” RNA

--Key points 7-4 “Seeing” RNA

--Video 7-4 “Seeing” RNA

--Discussion 7-4 “Seeing” RNA

-C7-Lecture 4-Homework

-Chapter 7 Test

-Learning Resources

--References

Chapter 8 Protein Synthesis, translation

-Lecture 1 Genetic codons and ribosome

--Key points 8-1 Genetic codons and ribosome

--Video 8-1 Genetic codons and ribosome

--Discussion 8-1 Genetic codons and ribosome

--C8-Lecture 1-Homework

-Lecture 2 Process of translation

--Key points 8-2 Process of translation

--Video 8-2 Process of translation

--Discussion 8-2 Process of translation

--C8-Lecture 2-Homework

-Lecture 3 Regulation of gene expression at Translational level

--Key points 8-3 Regulation of gene expression at Translational level

--Video 8-3 Regulation of gene expression at Translational level

--Discussion 8-3 Regulation of gene expression at Translational level

--C8-Lecture 3-Homework

-Lecture 4 Looking for your protein

--Key points 8-4 Looking for your protein

--Video 8-4 Looking for your protein

-Chapter 8 Test

-Learning resources

--References

--Life Cycle of an mRNA

--mRNA procesiing

--Protein synthesis

--SDS-PAGE

Final Exam

-Final Exam

Key points 4-5 Changing the natural gene sequences笔记与讨论

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