[农学]南京农业大学基因工程.ppt

Reporter genes Fig 11.16 半乳糖苷酶，新霉素磷酸转移酶，氯霉素乙酰转移酶，二氢叶酸还原酶，潮霉素磷酸转移酶，荧光素酶，葡糖醛酸酶 Fig 11.17 Deletion analysis. A reporter gene has been attached to the upstream region of a seed-specific gene from a plant. Deletion analysis -232 ~ -1 -617 ~ -1 -1078 ~ -1 GUS GUS GUS GhNP1 GhNP3 GhNP2 GhNP1 GhNP2 11.1 Studying the transcript of a cloned gene 11.2 Studying the regulation of gene expression 11.3 Identifying and studying the translation product of a cloned gene 11.3.1 HRT (Hybrid-release translation) and HART (hybrid-arrest translation) can identify the translation product of a cloned gene 11.3.2 Analysis of proteins by in vitro mutagenesis Cell-free translation Germinating wheat seeds or rabbit reticulocyte Fig 11.18 * Chapter 10. Studying gene location and structure Chapter 11. Studying gene expression and function Chapter 12. Studying genomics and post genomics Chapter 13. Production of protein from cloned genes Chapter 14 . Gene cloning and DNA analysis in medicines Chapter 15 . Gene cloning and DNA analysis in agriculture Chapter 16 . Gene cloning and DNA analysis in forensic science Part Two and Three The Applications of Gene Cloning and DNA Analysis in Research and Biotechnology Studying gene expression And function All genes have to be expressed in order to function. The first step in expression is transcription of the gene into a complementary RNA strand. To understand how a gene is expressed, the RNA transcript must be studied. ● ● Three type of transcription and translation Figure 11.1 exons Figure 11.1 5’ -UTR 3’ -UTR Figure 11.1 11.1.1 Detecting the presence of a Transcript and determining its nucleotide sequence Northern hybridization 11.1.1 Detecting the presence of a Transcript and determining its nucleotide sequence Northern hybridization R S L 4 8 12 16 20 24 28 32 36 40 44 48h Darkness 11.1.2 Transcript mapping by hybridization between gene and R