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时间:2021-10-19 17:56 来源: Essay代写

摘要:基因是DNA的片段,其提供了用于制备蛋白质的指示。这种蛋白质赋予生物体的特定性状或表型。遗传转化的字面意思......

Objective overview客观的概述

1.Perform genetic transformation through the method of bacterial transformation通过细菌转换的方法进行遗传转化

2.Observe the results of bacterial transformation in various growth condition and determine the degree of success of your efforts to genetically alter an organism.观察细菌转换的结果在不同的生长条件和确定你的努力的程度的成功基因改变有机体

Before coming to the lab

1.Read  section 12.6, 13.1, 14.2, 18.2 and 18.4 table in “Life – The Science of Biology”, 9th edition”; Sadava, Hillis, Heller, Berenbaum, 

2.Describe the flow of genetic material

3.Understand the transformation protocol

Introduction

A gene is a fragment of DNA which provides the instructions for making a protein. This protein gives an organism a particular trait or phenotype. Genetic transformation literally means change caused by genes, and involves the insertion of a gene into an organism in order to change the organism's trait. In this lab you will perform genetic transformation to E. coli. This is a very rare event, in which only about 1 in a million (106) cells take up the DNA.  Genetic transformation is used in many areas of biotechnology. In agriculture, genes coding for traits such as resistance to frost, pest, or spoilage can be genetically transformed into plants. In bioremediation, bacteria can be transformed with genes enabling them to digest oil spills. In medicine, diseases caused by defective genes are beginning to be treated by gene therapy; that is, by genetically transforming a sick person's cells with functioning copies of the defective gene that causes the disease.

 

You will use a procedure to transform bacteria with a gene that codes for Green Fluorescent Protein (GFP). The real-life source of this gene is the jellyfish Aequorea victoria. Green Fluorescent Protein causes the jellyfish to fluoresce around the rim of the organism (for unknown reasons). Following the transformation procedure, the bacteria express their newly acquired jellyfish gene and produce the fluorescent protein, which causes them to glow a brilliant green color when the GFP is excited by ultraviolet light. 

 

In this activity, you will learn about the process of moving genes from one organism to another with the aid of a plasmid. The method you are using today to transfer the plasmid to the bacteria is called bacterial transformation. It is the process in which bacteria take up free DNA in the environment. In this lab, you will be forcing Escherichia coli cells to take up DNA, but this process occurs spontaneously in nature, with some bacteria, leading to bacteria acquiring new traits, such as antibiotic resistance.   This process, called horizontal gene transfer, is the bacterial version of the recombination of genes that normally occurs during sex in higher organisms.  Because of this horizontal gene transfer in bacteria, it is more difficult to determine the species of a bacterial cell, as well as determine the evolutionary relatedness of bacteria.  This lab has important links between the two modules of this course:  “Is there life out there?” and “Why Have Sex?”.  You will be considering these links in the second week of this lab.

 

Bacteria often conduct transformation using small circular pieces of DNA call plasmids that are separate from their much larger chromosomes. Plasmid DNA usually contains genes for one or more traits that may be beneficial to bacterial survival. In nature, bacteria can transfer plasmids back and forth allowing them to share these beneficial genes. This natural mechanism allows bacteria to adapt to new environments. A notable example of this adaptation is the occurrence of bacterial resistance to antibiotics, which is due, in part, to the transmission of plasmids. 

 

pGLO plasmid carries the gene encoding GFP and a gene for resistance to the antibiotic ampicillin. pGLO also incorporates a special gene regulation system, which can be used to control transcription (expression) of the fluorescent protein in transformed cells. The gene for GFP can be switched “on” (transcribed) in transformed cells by adding the sugar arabinose to the cells' nutrient medium.  Arabinose binds to the activator protein, AraC, and allows the AraC activator to bind to DNA near the promoter of GFP genes.  This allows the RNA polymerase to bind the promoter and start transcribing the GFP genes.  When arabinose is absent, the AraC activator cannot bind to the proper location on the DNA to activate expression (transcription) of the GFP genes.   

Pre-lab Questions

1. To genetically transform an entire multicellular organism, you must insert the new gene into every cell in the organism. Which organism is more easily transformed: a multicellular organism or a single-celled organism?

本文标签: 细菌转化

[出国申请的essay要怎么写]细菌转化Bacterial transformationhttps://www.0592w.com/ESSAY/16342.html

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