Buccal Dna Collection Using Treated Paper

This study presents a domestic kit suitable for the collection of human buccal cells, and long-term storage of nucleic acids on a piece of paper. A filter paper with a density of 70 g/m2 was selected and treated with chemicals that make up the lysing solution, which contributes to the destruction of cells and prevents bacterial growth. A total of 25 buccal cell samples were collected on the treated paper cards. The samples had a median DNA concentration of 20 ng/μl (range 0.5–45 ng/μl). The collected DNA samples were tested using a real-time PCR, sequencing of the first hypervariable segment of the human mtDNA control region, and multiplex genotyping of 15 short tandem repeat loci. We conclude that the domestic pretreated cards could be used for collecting and isolating DNA from buccal epithelium for genetic and other applications.

ID: 1049 Variation of mitochondrial DNA HV1 and HV2 of the Vietnamese population

The sequence polymorphism of mitochondrial DNA (mtDNA) hypervariable Segment 1 (HV1) and hypervariable Segment 2 (HV2) are studied and applied to genetic diversity and human evolution assessment, forensic genetics, consanguinity determination, and mitochondrial disease diagnosis. Nucleotide sequence variations in HV1 and HV2, two hypervariable segments of the noncoding control region of human mitochondrial DNA (mtDNA), in selected ethnics of the Vietnamese population were elucidated through sequencing. In this study, we define the variations of HV1 and HV2 of 517 unrelated Vietnamese individuals in Kinh, Muong, Cham, and Khmer ethnic. We found 50 haplogroups: F1a haplogroup frequency is the highest at 15.7%; B5a haplogroup frequency is 10.8%, M haplogroup frequency is 8.9%, M7b1 haplogroup frequency is 7.7%; B6, D4e, D5a, E, F1c, F2a, F3a, G2a, M9b, N, N21 and U5a haplogroup frequencies are the lowest (1%). The frequency of SNP A263G are 100%; A73G is 99.6%, 315insC is 96%; 309insC is 56%; C16223T is 41%, and T16189C is 39%. We have assessed the genetic polymorphism of mtDNA HV1 and HV2 of 517 Kinh, Muong, Cham, Khmer ethnic samples.

DIVERSITAS GENETIK DAN HAPLOGROUP KAMBING GEMBRONG BERSTATUS KRITIS DI KABUPATEN KARANGASEM, BALI (Genetic Diversity and Haplogroup of Endangered Gembrong Goat In Karangasem, Bali)

This study was conducted to evaluate the genetic diversity and phylogeny of Gembrong goat. For this purpose, 21 goats from endangered breed in Karangasem Bali were used. Molecular analysis of genetic diversity and phylogeography used hypervariable segment 1 of mitochondrial DNA control region. The result showed that genetic variability of Gembrong goat was homogeneous with only one different sites, namely the substitution pyrimidines of C ↔ T (transitional). Phylogeny analysis results showed maternal origin of Gembrong goat is lineage (subhaplogroup) B1 with frequency of 100%. Haplogroup B were known has been domesticated from wild goat in western Asia, then headed to south Asia and infiltrated to southeast Asia, including Gembrong goat in Bali, Indonesia. As a conclusion, genetic diversity of Gembrong goat from remaining population in Karangasem very low and originate from lineages/haplogroup B1 with a frequency of 100%.

Localization of the Escherichia coli RNA Polymerase β′ Subunit Residue Phosphorylated by Bacteriophage T7 Kinase Gp0.7

ABSTRACT During bacteriophage T7 infection, the Escherichia coli RNA polymerase β′ subunit is phosphorylated by the phage-encoded kinase Gp0.7. Here, we used proteolytic degradation and mutational analysis to localize the phosphorylation site to a single amino acid, Thr1068, in the evolutionarily hypervariable segment of β′. Using a phosphomimetic substitution of Thr1068, we show that phosphorylation of β′ leads to increased ρ-dependent transcription termination, which may help to switch from host to viral RNA polymerase transcription during phage development.