Inventors: Assist. Prof. Dr. Faiz Fayyad Mohammed, Assist. Prof. Dr. Zainab Karim Kazim/Department of Field Crops, College of Agricultural Engineering Sciences, University of Baghdad
Abstract
Genetic mutations are alterations caused by the insertion, deletion or substitution of the nitrogenous bases of a gene sequence, leading to a change in the structure of the gene and consequently an alteration in the structure of the protein resulting in a change of the phynotype. There are many Kits that are produced by different companies and that can be used to create genetic mutations at the gene level, but these kits are characterized by their too many components, steps, high prices and the need for precision in carrying out their steps, as well as these kits require a high-efficient bacteria (Super competent Escherichia coli cells) that equipped by specialized companies for that purpose.
Also, the use of these kits may sometimes not succeed to induce genetic mutation. So this study aimed to find a simple and inexpensive method to induce a genetic mutation in one of the P3 genes of potato virus Y (PVY). The new method is designed with four primers, two of which contain the target genetic mutation (insertion, deletion or substitution of nitrogenous bases in the gene) with three polymerase chain reactions (PCR). This method was characterized by very high accuracy, applicability and effort and time saving. The new method cost no more than $100 compared to the kits supplied by the companies, which may cost more than $1000. Furthermore, with the new method, it is possible to use the normal competent E. coli cells instead of the supercompetent E. coli cells which is extremely required to be used with the kit supplied by the manufacturer.