Bacteria of the genus Mycoplasma have among the smallest known genomes for a free-living cell (only 525 genes!). As we discussed in class, a Mycoplasma also served as the basis of the first completely "synthetic" organism. You are tasked with trying to create a truly minimal version of this microbe - in other words, a version containing only the genes absolutely essential for life. a) Many different Mycoplasma genomes are available in reference databases. Briefly describe an analysis you could conduct with these other genomes to help inform you which genes might be essential, summarizing what you would look for and how this would relate to the question of likely essential genes. b) To complement this bioinformatic approach, briefly outline a genetic screen that would allow you to identify essential protein-coding genes (ie we are looking for gene presence/absence, ignoring mutations influencing regulation). Assume any standard mutagenesis approach will work in this organism (chemical mutagens, transposons, etc). Detail how you will make your mutations, what phenotype you will look for in your mutant library, and how you will identify where your mutations occurred. c) To test your minimal microbe, your final job is to artificially synthesize this new genome from scratch. Briefly, why must you do this from small pieces, rather than simply synthesizing the entire DNA molecule at once?
Molecular Techniques
Molecular techniques are methods employed in molecular biology, genetics, biochemistry, and biophysics to manipulate and analyze nucleic acids (deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)), protein, and lipids. Techniques in molecular biology are employed to investigate the molecular basis for biological activity. These techniques are used to analyze cellular properties, structures, and chemical reactions, with a focus on how certain molecules regulate cellular reactions and growth.
DNA Fingerprinting and Gel Electrophoresis
The genetic makeup of living organisms is shown by a technique known as DNA fingerprinting. The difference is the satellite region of DNA is shown by this process. Alex Jeffreys has invented the process of DNA fingerprinting in 1985. Any biological samples such as blood, hair, saliva, semen can be used for DNA fingerprinting. DNA fingerprinting is also known as DNA profiling or molecular fingerprinting.
Molecular Markers
A known DNA sequence or gene sequence is present on a chromosome, and it is associated with a specific trait or character. It is mainly used as a genetic marker of the molecular marker. The first genetic map was done in a fruit fly, using genes as the first marker. In two categories, molecular markers are classified, classical marker and a DNA marker. A molecular marker is also known as a genetic marker.
DNA Sequencing
The most important feature of DNA (deoxyribonucleic acid) molecules are nucleotide sequences and the identification of genes and their activities. This the reason why scientists have been working to determine the sequences of pieces of DNA covered under the genomic field. The primary objective of the Human Genome Project was to determine the nucleotide sequence of the entire human nuclear genome. DNA sequencing selectively eliminates the introns leading to only exome sequencing that allows proteins coding.
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