Lab Report About Lab
Abstract In food industry such as the production of cheese, bacteria takes a significant role on the wide variety of cheese. Lactic acid bacteria(LAB), a bacteria that can be found in the production of cheese, its stress gene was investigated in the experiment by using various biochemical and genetic techniques to identify and extract. The characterisation of the strain illustrates how identification of strains differ using different methods, such as gram stain and 16s rRNA screening. After the characterisation, the stress gene isolation assist the further understanding of the gene on LAB be giving different stress in future work.
Aims
Whole experiment can be separated into two parts
(I) The characterisation of the isolate(lactococcal species) given by * Using simple biochemical tests * Using genetic tests by extraction of total genomic DNA * Amplifying the gene encoding for the 16s rRNA by PCR * Understanding the fermentation growth kinetics in controlled liquid media to know the capability of organisms * The investigation of the digestion of the 16s rDNA amplicon with Hinf1 on 16s rDNA strains
(II) The amplification and cloning of a specific stress gene given by * Designing primers for cloning * cloning stress gene into cloning vector (pGEM-T) * cloning stress gene into expression plasmid(pET23b) – not done
Literature Review
Lactic acid bacteria (LAB)
LAB are defined as bacteria that produce lactic acid as their major fermentation production. The examples of LAB include Streptococcus, Enterococcus, Leuconostoc and Lactococcus. They are gram-positive rods or cocci bacteria with high AT content. (Prescott et. al, 2008). There are few applications of LAB in food, medical and biotechnological industries. The major use of LAB is on the food industries. The non-pathogenic species, which helps the process of fermentation of different food such as dairy products, meat and vegetables. They mainly convert the hexose
Aims
Whole experiment can be separated into two parts
(I) The characterisation of the isolate(lactococcal species) given by * Using simple biochemical tests * Using genetic tests by extraction of total genomic DNA * Amplifying the gene encoding for the 16s rRNA by PCR * Understanding the fermentation growth kinetics in controlled liquid media to know the capability of organisms * The investigation of the digestion of the 16s rDNA amplicon with Hinf1 on 16s rDNA strains
(II) The amplification and cloning of a specific stress gene given by * Designing primers for cloning * cloning stress gene into cloning vector (pGEM-T) * cloning stress gene into expression plasmid(pET23b) – not done
Literature Review
Lactic acid bacteria (LAB)
LAB are defined as bacteria that produce lactic acid as their major fermentation production. The examples of LAB include Streptococcus, Enterococcus, Leuconostoc and Lactococcus. They are gram-positive rods or cocci bacteria with high AT content. (Prescott et. al, 2008). There are few applications of LAB in food, medical and biotechnological industries. The major use of LAB is on the food industries. The non-pathogenic species, which helps the process of fermentation of different food such as dairy products, meat and vegetables. They mainly convert the hexose