Cloning Of Il2 In Ptz19r And Its Site Directed Mutagenesis Using Kunkel’s Method

HARIPRIYA., .N and N, M., SIVANANDHAM (2014) Cloning Of Il2 In Ptz19r And Its Site Directed Mutagenesis Using Kunkel’s Method. In: International Conference on Advances In Engineering And Technology - ICAET 2014, 24 - 25 May, 2014, RIT, Roorkee, India.

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Abstract- Interleukin-2 (IL-2) is a lymphokine that exerts immunoregulatory effects on a variety of cells such as T cells, activated B cells and Natural Killer (NK) cells. Its biological effects are mediated through specific interactions with cell surface receptors present on target cells. Due to its role in generating normal immune responses, IL 2 is used in the treatment of a variety of tumors and infectious diseases. The downside of IL 2 is that the molecule is rather unstable. The aim of the current research is to generate IL-2 mutant with improved potency.This objective is accomplished by the use of cloning and Site directed Mutagenesis (SDM). For this purpose,first, wild type IL-2 mRNA was isolated from cultured T lymphocytes. This was converted into cDNA with the help of Reverse Transcription-PCR. Future work includes amplification of cDNA fragments and cloning into a phagemid vector pTZ19R. Transformation will be carried out in E.coli RZ1032 and the ssDNA obtained as the end result of phagemid replication will be used as a template DNA to carry out modified Kunkel’s method of site directed mutagenesis.

Item Type: Conference or Workshop Item (Paper)
Uncontrolled Keywords: words: lymphokine, Reverse Transcription PCR, phagemid vector,Transformation
Depositing User: Mr. John Steve
Date Deposited: 27 May 2019 04:43
Last Modified: 27 May 2019 04:43

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