Sodium Cloride And Sucrose Rescued Cupriavidus Necator H16 Δsecb Mutant When Grown In Nutrient Rich Media
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...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2016
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| Subjects: | |
| Online Access: | http://eprints.usm.my/31855/1/SIM_XUAN_YI_24%28NN%29.pdf http://eprints.usm.my/31855/ |
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| Summary: | 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.
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