A study of degraded skeletal samples using ForenSeq DNA Signature™ Kit

Recent advances in massively parallel sequencing (MPS) has become a very promising technology for massive genetic sequencing [1]. In this study Illumina ForenSeq™ DNA Signature Prep Kit was tested to determine if MPS offers a more comprehensive evaluation of degraded samples than the traditional fragment analysis/capillary electrophoresis based method. The Illumina® ForenSeq™ DNA Signature MPS Kit, includes 200 genetic loci [2]. The use of NGS would therefore reduce the analysis time and augment the identification of human remains. In this context we aimed to analyse the hard tissue degraded samples using Illumina® ForenSeq™ DNA Signature MPS Kit. These samples had given partial profiles with dropout at several loci with GlobalFiler™ kit previously. The MPS kit showed that it is highly sensitive, aids in higher allele recovery for STR loci and provides valuable information about biogeographic ancestry, identity and phenotypic features from a single analysis. The work resulted in highly successful amplification and sequencing of 30 degraded bone/teeth samples using MPS method

A novel 3-hydroxypropionic acid-inducible promoter regulated by the LysR-type transcriptional activator protein MmsR of Pseudomonas denitrificans

MmsR (33.3 kDa) is a putative LysR-type transcriptional activator of Pseudomonas denitrificans. With the help of 3-hydroxypropionic acid (3-HP), an important platform chemical, MmsR positively regulates the expression of mmsA, which encodes methylmalonylsemialdehyde dehydrogenase, the enzyme involved in valine degradation. In the present study, the cellular function of MmsR and its binding to the regulatory DNA sequence of mmsA expression were investigated both in vivo and in vitro. Transcription of the mmsA was enhanced >140-fold in the presence of 3-HP. In the MmsR-responsive promoter region, two operators showing dyad symmetry, designated O-1 and O-2 and centered at the -79 and -28 positions, respectively, were present upstream of the mmsA transcription start site. An electrophoretic mobility shift assay indicated that MmsR binds to both operator sites for transcription activation, probably in cooperative manner. When either O-1 or O-2 or both regions were mutated, the inducibility by the MmsR-3-HP complex was significantly reduced or completely removed, indicating that both sites are required for transcription activation. A 3-HP sensor was developed by connecting the activation of MmsR to a green fluorescent readout. A more than 50-fold induction by 25 mM 3-HP was observed