Sodium Cloride And Sucrose Rescued Cupriavidus Necator H16 Δsecb Mutant When Grown In Nutrient Rich Media

• Main Author: Sim , Xuan Yi
• Format: Thesis
• Language: English
• Published: 2016
• Subjects: QH1 Natural history (General - Including nature conservation, geographical distribution)
• Online Access: http://eprints.usm.my/31855/1/SIM_XUAN_YI_24%28NN%29.pdf; http://eprints.usm.my/31855/
• Institution: Universiti Sains Malaysia

SecB adalah salah satu komponen utama dalam sistem translokasi protein Sec terutamanya bagi bakteria Gram-negatif. Dalam kajian ini, fungsi SecB telah dikaji dengan membina mutan Cupriavidus necator H16 ΔsecB. Mutan ini telah dibina melalui pertukaran alel menggunakan vektor swa-hapus pDM4. Untuk mengesahkan perubahan fenotip disebabkan oleh gen yang dimutasi, mutan pelengkap juga dibina, yang mana vektor pBBR1MCS2 yang mengandungi gen SecB telah dikonjugasi ke dalam mutan C. necator H16 ΔsecB. Jenis liar, C. necator H16 ΔsecB mutan dan mutan pelengkap telah dikaji mengguna sistem Biolog Phenotype Microarray. Mutan pelengkap menunjukkan fenotip jenis liar, bermakna penglengkapan secB telah berjaya. Mutan Cupriavidus necator H16 ΔsecB tidak menunjukkan perbezaan yang signifikan apabila ditumbuh dialam kebanyakan substrat. SecB is one of the key players in the Sec protein-translocation system ubiquitous to Gram-negative bacteria. In this study the function of SecB was studied by studying an unmarked Cupriavidus necator H16 ΔsecB mutant. The mutant was constructed through allelic exchange utilizing the pDM4 suicidal vector. In order to confirm that the phenotypic change is due to the mutated gene, a complementation mutant was also constructed, in which vector pBBR1MCS2 harbouring a secB gene was introduced into the C. necator H16 ΔsecB mutant via conjugation. The wild type, C. necator H16 ΔsecB mutant and the complementation mutant were characterised using Biolog Phenotype Microarray. The result shows that the complementation mutant exhibits the wild type phenotype, indicating successful complementation of the unmarked secB deletion. The C. necator H16 ΔsecB mutant did not show any significant difference when grown on most substrate.