Molecular characterization of the Philippine carabao (Bubalus bubalis L.) from the major island groups of the Philippines using molecular cloning and sequence analysis of the D-loop of mitochondrial DNA

Date

4-2009

Degree

Bachelor of Science in Biology

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Cella B. Dela Viña

Co-adviser

Medino Gedeun N. Yebron Jr.

Abstract

The mtDNA D-loop sequences were used to characterize the Philippine carabao (Bubalus bubalis L.) at the molecular level using molecular cloning and sequence analysis. A total of 36 blood and DNA samples from Philippine carabaos were collected from the major island groups of the Philippines specifically in Cagayan, Batangas, Bohol and Bukidnon. The D-loop region was amplified through PCR primers CB3 and A.dLo.H with PCR product of 484-bp. PCR products were cloned, and sent for sequencing. Sequence analysis of all the Philippine carabao isolates presented a 97.5% homology. This showed that the D-loop sequences from the major island groups of the Philippines are highly homologous. BLAST-N analysis was done on representative sequences and results showed that the isolate from sample 1 from Batangas, A2 from Bukidnon, Cl and C9 from Cagayan are 99-100% similar to the FuLing1213 isolate from China. Likewise, sample 5 from Batangas and Al from Bukidnon are similar with the Xinglong isolate of China, sample B1 from Bohol with the Dechang isolate of China and sample BIO from Bohol with the Enshi isolate of China. This implies that the Philippine carabao descended from these maternal lines. Characterization of the D-loop sequence obtained from the Philippine carabao showed that the sequenced region was part of the ETAS and central domain. A rooted phylogenetic tree confirmed that it belongs to Lineage A of swamp buffaloes and that the Philippine carabao is closely related to the Chinese buffaloes. The clones obtained from this study may be helpful in the development of species identification methods.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

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