Welcome Note from Editor-in-Chief

It is with great pleasure that I write this editorial to welcome you to the first issue of this new International journal, “Pakistan Biomedical Journal” (PBMJ). The topics covered by the journal are certainly broad and interesting. Biomedical science is a collection of applied sciences that help us understand, research, and innovate within the field of healthcare. It includes disciplines like molecular biology, clinical virology, bioinformatics, and biomedical engineering, among others. It's designed to apply the biological sciences to advance not only individual health but also the area of public health. Biomedical Research can help health professions better understand things like the human body and cell biology, making advances in our understanding of epidemics, health initiatives, and human health in the age of longer life expectancy. It aids our understanding of infectious disease and provides research opportunities into some of our most troubling health issues. The journal will continue to publish high quality clinical and biomedical research in health and disease later in life. Peer review will remain a vital component of our assessment of submitted articles.I am very happy to have a team of excellent editors and editorial board members from the top international league covering in depth the related topics. They will ensure the highest standards of quality for the published manuscripts and, at the same time, keep the process time as short as possible. We hope to bring best researches in the field of biomedical sciences that may serve as a guideline in health awareness, understanding the mechanisms and its management in future.   We definitely look forward to receiving your excellent studies to making PBMJ synonymous with high quality in the biomedical science domain.

Expression and Transformation Studies of Antifreeze Protein Gene of Carrot in Potato

The Antifreeze protein gene (1.022Kb) from local of D.carrota cultivar T29 was amplified from four weeks old seedling leave tissue genomic DNA. The PCR amplified gene was cloned in TA-Cloning vector pCR2.1. The cloned DcAFP gene was then sequenced, the DNA BLAST homology with wild carrot AFP was 96.1%. The 999bp gene of DcAFP from T29 cultivar was translated into amino acid sequence, the translated amino acid sequence when aligned with wild carrot AFP gene of amino acid sequence. The gene has 13 amino acid variability. The Asparagine amino acid residues which play an important role as an ice interacting/ice binding sites were intact as in wild species of carrot amino acid sequence, but extra three Asparagine amino acid were noted in 332 amino acid gene sequence as in the case of DcAFP (T29). The 944bp DNA sequence that code the mature peptide of DcAFP was cloned in pET15b E.coli expression vector. The IPTG induced 36kDa protein detected from the BL21 expression host transformed with expression plasmid. The 36kDa recombinant protein inclusion refolded and purified for polyclonal antibody production. Purified IgG of DcAFP was used in western blot, protein dot blot and ELISA analysis of transgenic potato lines. The 999bp full length gene was ligated in pCAMBIA-1301 without selection marker. Plant expression construct pCAMBIA1301hph - DcAFP (T29) was transformed in Agrobacterium host strain LBA4404. Agrobacterium with pCAMBIA construct was transformed in potato nodal explant. The transgenic plant first selected by PCR, DNA dot blot and GUS reporter assay. Stable integration confirmed by southern blot analysis. The DcAFP mRNA detected by Northern blotting after cold induction of transgenic potato plants. DcAFP protein detected by protein dot blot and western blotting methodologies. The sandwich ELISA was successfully done for expressed protein estimation in transgenic potato lines. The stable potato transgenic lines were tested with ILA (Ion Leakage Assay), which results are significant when compared with the control plants. It was noted that due to DcAFP protein expression in transgenic potato lines demonstrating that transgenic plants are tolerating the frost temperature.