Publication: Cloning of pre-mir-367 coding sequence by homologous recombination and DNA ligation methods
| dc.contributor.author | Wun, Lim Yin | |
| dc.date.accessioned | 2026-09-20T07:38:16Z | |
| dc.date.available | 2026-09-20T07:38:16Z | |
| dc.date.issued | 2026-01 | |
| dc.description.abstract | MicroRNAs (miRNAs) are important post-transcriptional regulators involved in diverse biological processes. Pre-miRNAs serve as essential intermediates in miRNA biogenesis, and the construction of pre-miRNA expression vectors is critical for functional characterisation of specific miRNAs such as pre-miR-367. Plasmid construction in our laboratory has primarily been performed using restriction-ligation cloning, while homologous recombination has not yet been applied for assembling plasmid containing microRNA inserts. This study aimed to construct a recombinant pCMV-MiR plasmid carrying the pre-miR-367 sequence using two cloning strategies: homologous recombination and restriction–ligation cloning. Cloning outcomes were verified using colony PCR, agarose gel electrophoresis, and Sanger sequencing. Homologous recombination produced bacterial colonies, with initial colony PCR indicating potential recombinant clones; however, Sanger sequencing confirmed the absence of the pre-miR-367 insert in the recovered plasmids. In contrast, the restriction–ligation approach did not yield transformant colonies under the conditions tested. Although a sequence-verified recombinant pCMV-MiR–pre-miR-367 construct was not obtained, this study highlights key technical challenges associated with cloning small and structurally complex pre-miRNA inserts and provides methodological insights that may support future optimisation of miRNA expression vector construction. | |
| dc.identifier.uri | https://erepo.usm.my/handle/123456789/25038 | |
| dc.language.iso | en | |
| dc.title | Cloning of pre-mir-367 coding sequence by homologous recombination and DNA ligation methods | |
| dc.type | Resource Types::text::thesis::bachelor thesis | |
| dspace.entity.type | Publication | |
| oairecerif.author.affiliation | Universiti Sains Malaysia |