Arrange the steps that would be used in a laboratory to engineer a bacterium that could express the human gene coding for factor VIII. Not all steps will be placed. Incorrect Isolate the human gene that produces factor VIII. Isolate the mRNA of the factor VIII gene. Generate cDNA of the factor VIII gene using reverse transcriptase. Insert the factor VIII cDNA into a bacterial vector near a promoter site. Transform the vector into an E. coli bacterium. Human DNA is introduced into the bacterial cell using direct injection by a small needle. E. coli expresses factor VIII. Answer Bank The factor VIII protein is isolated from human tissues.
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.
Molecular cloning is used to transfer a foreign DNA into the host (bacteria) genome that is responsible for the expression of the foreign DNA in the host cells. A vector is used to transfer the DNA from the foreign organism to the host organism.
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