Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development

The Newcastle disease virus can replicate rapidly and kill human cancer cells. Therefore it has the potential to be developed as a cancer vaccine. However, the immune system hinders the replication of the virus in these cells. In order to make it more efficient as a cancer vaccine, the the human int...

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Main Author: Che Ani, Muhamad Alhapis
Format: Project Paper Report
Language:English
Published: 2015
Online Access:http://psasir.upm.edu.my/id/eprint/85123/
http://psasir.upm.edu.my/id/eprint/85123/1/FBSB%202015%20112%20-%20IR.pdf
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author Che Ani, Muhamad Alhapis
author_facet Che Ani, Muhamad Alhapis
author_sort Che Ani, Muhamad Alhapis
building UPM Institutional Repository
collection Online Access
description The Newcastle disease virus can replicate rapidly and kill human cancer cells. Therefore it has the potential to be developed as a cancer vaccine. However, the immune system hinders the replication of the virus in these cells. In order to make it more efficient as a cancer vaccine, the the human interleukin-12 (hIL-12) gene will be cloned into the virus. However, before this gene can be transferred to the virus, it must first be cloned into a cloning vector. In this project, the hIL-12 gene was obtained from the plasmid pUNO-hIL12 and cloned into a pJET cloning vector. A set of forward and reverse primers with NheI restriction included was designed to amplify hIL-12 gene. This will lead to producing the hIL-12 gene fragment with NheI restriction sites after amplification by polymerase chain reaction (PCR) cycles. The PCR product was then purified by using MEGAquick-spinTM Total Fragment DNA Purification Kit. The purified hIL-12 was then cloned into pJET cloning vector by T4 DNA ligase and transformed into competent Escherichia. coli Top 10 cells by using heat-shock transformation method. The cloning was verified using NheI restriction enzyme digestion analysis and further confirmed by sequencing.
first_indexed 2025-11-15T12:38:16Z
format Project Paper Report
id upm-85123
institution Universiti Putra Malaysia
institution_category Local University
language English
last_indexed 2025-11-15T12:38:16Z
publishDate 2015
recordtype eprints
repository_type Digital Repository
spelling upm-851232021-12-27T07:55:23Z http://psasir.upm.edu.my/id/eprint/85123/ Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development Che Ani, Muhamad Alhapis The Newcastle disease virus can replicate rapidly and kill human cancer cells. Therefore it has the potential to be developed as a cancer vaccine. However, the immune system hinders the replication of the virus in these cells. In order to make it more efficient as a cancer vaccine, the the human interleukin-12 (hIL-12) gene will be cloned into the virus. However, before this gene can be transferred to the virus, it must first be cloned into a cloning vector. In this project, the hIL-12 gene was obtained from the plasmid pUNO-hIL12 and cloned into a pJET cloning vector. A set of forward and reverse primers with NheI restriction included was designed to amplify hIL-12 gene. This will lead to producing the hIL-12 gene fragment with NheI restriction sites after amplification by polymerase chain reaction (PCR) cycles. The PCR product was then purified by using MEGAquick-spinTM Total Fragment DNA Purification Kit. The purified hIL-12 was then cloned into pJET cloning vector by T4 DNA ligase and transformed into competent Escherichia. coli Top 10 cells by using heat-shock transformation method. The cloning was verified using NheI restriction enzyme digestion analysis and further confirmed by sequencing. 2015 Project Paper Report NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/85123/1/FBSB%202015%20112%20-%20IR.pdf Che Ani, Muhamad Alhapis (2015) Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development. [Project Paper Report]
spellingShingle Che Ani, Muhamad Alhapis
Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development
title Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development
title_full Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development
title_fullStr Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development
title_full_unstemmed Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development
title_short Cloning of human- interleukin-12 into pJET cloning vector for anti-cancer vaccine development
title_sort cloning of human- interleukin-12 into pjet cloning vector for anti-cancer vaccine development
url http://psasir.upm.edu.my/id/eprint/85123/
http://psasir.upm.edu.my/id/eprint/85123/1/FBSB%202015%20112%20-%20IR.pdf