Journal of Genetic Engineering and Biotechnology

Exopolysaccharides (EPSs) từ vi sinh vật là các biopolymer tự nhiên vô hại quan trọng, được sử dụng trong các ứng dụng bao gồm dược phẩm, nutraceuticals và thực phẩm chức năng, mỹ phẩm và thuốc trừ sâu. Nhiều loại vi sinh vật có khả năng tổng hợp và bài tiết EPSs với các tính chất hóa học và cấu trúc khiến chúng phù hợp với một số ứng dụng quan trọng. Vi sinh vật bài tiết EPS ra bên ngoài màng tế bào của chúng, dưới dạng chất nhờn hoặc “thạch” vào môi trường ngoại bào. Những vi sinh vật sản xuất EPS này rất phổ biến và có thể được phân lập từ các môi trường nước và đất, chẳng hạn như nước ngọt, nước biển, nước thải và đất. Chúng cũng đã được phân lập từ những môi trường cực đoan như suối nước nóng, nước lạnh, các môi trường ưa muối và đầm lầy muối. Gần đây, EPS vi sinh vật đã thu hút sự quan tâm cho các ứng dụng như tác nhân kết tụ sinh học môi trường nhờ tính chất phân hủy được và không độc hại của chúng. Tuy nhiên, cần có thêm nỗ lực để sản xuất EPS vi sinh vật một cách hiệu quả về chi phí và quy mô công nghiệp. Bài đánh giá này tập trung vào các exopolysaccharides thu được từ nhiều vi sinh vật ưa cực, quá trình tổng hợp của chúng và tối ưu hóa sản xuất để cải thiện chi phí và năng suất. Chúng tôi cũng đã khám phá vai trò và ứng dụng của chúng trong các tương tác giữa nhiều sinh vật khác nhau.

Insulin-like growth factor 2 (IGF2) is one of three hormones that share high structural similarity to insulin. It is involved in several insulin-like growth-regulating and mitogenic activities. This study was conducted to genotype the coding regions of the IGF2 gene in Japanese quail (Coturnix japonica) using PCR-SSCP-sequencing, and to assess the possible association of the polymorphism of these regions with the main egg production traits. A total of 240 female birds with an equal number of three Japanese quail populations (Brown or BR, Black or BL, and White or WT) were included in this study. All the genotyped regions exerted no heterogeneity in their sequences with one exception detected in the exon 2. In this locus, a novel single nucleotide polymorphism (SNP) was detected in which “A” was substituted with “G” at 81 position with a silent effect (p.F79=SNP) on IGF2 protein. Association analyses indicated a significant (P < 0.05) relation of this SNP with egg number (EN) and bird weight (BW) in the analyzed populations, in which the birds with AG genotype had lower EN and BW than those with AA genotype. The p.F79=SNP was largely detected in the WT line than the other two lines. The detected p.F79=SNP has a highly negative effect on EN and BW in Japanese quail. Thus, the implementation of the variations of the IGF2 gene can be a useful marker in the marker-assisted selection of Japanese quail. This is the first report to describe IGF2 gene variations in Japanese quail, which strongly suggests raising the birds from the BR line with AA genotype when breeders desire to raise Japanese quail for large-scale egg production.

In the current research, we have developed silver and iron nanoparticles of isolated proanthocynidin (PAC) from grape seed by green synthesis and evaluated for antimicrobial, antioxidant activity and in vitro cytotoxicity against colon cancer cell lines. One percent solution of isolated proanthocynidin in water was vigorously mixed with 1% silver nitrate and 1% ferric chloride solution and kept for 4 h, to yield PACAgNP and PACFeNP. The synthesized nanoparticles were characterized by UV, FTIR, XRD, and SEM analysis and evaluated for antimicrobial potential against selected microbes. Moreover, the synthesized nanoparticles were studied for DPPH assay and in vitro cytotoxicity using colon cancer cell lines COLO320DM and HT29 (MTT, SRB, and Trypan blue assay). UV spectroscopy confirmed the development of nanoparticles. SEM analysis showed that the particles were aggregated in the size range of 50 to 100 nm. Antimicrobial potential was found to be less against Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli, whereas cytotoxicity of PACAgNP and PACFeNP against COLO320DM and HT29 exhibited promising results as compared to the pure PAC. PACAgNP and PACFeNP exhibited 20.83 ± 0.33% and 18.06 ± 0.60% inhibition, respectively, against DPPH radical, whereas pure PAC showed 16.79 ± 0.32% inhibition and standard (ascorbic acid) exhibited 98.73 ± 0.18% inhibition of DPPH radical. The silver and iron nanoparticles were successfully developed by green synthesis method using isolated proanthocynidin which is economical and eco-friendly. The use of metal nanoparticles may open up a new opportunity for anticancer therapies to minimize the toxic effects of available anticancer drugs specifically in targeting specific site.

The Capparaceae family is commonly recognized as a caper, while Cleomaceae represents one of small flowering family within the order Brassicales. Earlier, Cleomaceae was included in the family Capparaceae; then, it was moved to a distinct family after DNA evidence. Variation in habits and a bewildering array of floral and fruit forms contributed to making Capparaceae a “trash-basket” family in which many unrelated plants were placed. Indeed, family Capparaceae and Cleomaceae are in clear need of more detailed systematic revision. Here, in the present study, the morphological characteristics and the ecological distribution as well as the genetic diversity analysis among the twelve species of both Capparaceae and Cleomaceae have been determined. The genetic analysis has been checked using 15 ISSR, 30 SRAP, and 18 ISTR to assess the systematic knots between the two families. In order to detect the molecular phylogeny, a comparative analysis of the three markers was performed based on the exposure of discriminating capacity, efficiency, and phylogenetic heatmap. Our results indicated that there is a morphological and ecological variation between the two families. Moreover, the molecular analysis confirmed that ISTR followed by SRAP markers has superior discriminating capacity for describing the genetic diversity and is able to simultaneously distinguish many polymorphic markers per reaction. Indeed, both the PCA and HCA data have drawn a successful annotation relationship in Capparaceae and Cleome species to evaluate whether the specific group sort individual or overlap groups. The outcomes of the morphological and ecological characterization along with the genetic diversity indicated an insight solution thorny interspecies in Cleome and Gynandropsis genera as a distinct family (Cleomaceae) and the other genera (Capparis, Cadaba, Boscia, and Maerua) as Capparaceae. Finally, we recommended further studies to elucidate the systematic position of Dipterygium glaucum.

Various Asparagus species constitute the significant vegetable and medicinal genetic resource throughout the world. Asparagus species serve as important commodity of food and pharmaceutical industries in India. A diverse collection of Asparagus species from different localities of Northwest India was investigated for its genetic diversity using simple sequence repeat (SSR) markers. Polymorphic SSR markers revealed high genetic diversity. Primer SSR-15 amplified maximum of 8 fragments while 3 primers, namely, SSR-43, SSR-63, and AGA1 amplified minimum of 3 fragments. Collectively, 122 alleles were amplified in a range between 3 and 8 with an average of 5 alleles per marker. The size of the amplified alleles ranged between 90 and 680 base pairs. Polymorphism information content (PIC) value varied from a highest value of 0.499 in primer AGA1 to a lowest value of 0.231 in primer SSR-63 with a mean value of 0.376 showing considerable SSR polymorphism. Dendrogram developed on the basis of Jaccard’s similarity coefficient and neighbor-joining tree segregated all the studied Asparagus species into two discrete groups. Structure analysis based on Bayesian clustering allocated different accessions to two independent clusters and exhibited low level of individual admixture. The genetic diversity analysis showed a conservative genetic background for maximum species of asparagus. Only Accessions of Asparagus adscendens were split into two diverse clusters suggesting a wide genetic base of this species as compared to other species. Overall genetic diversity was high, and this germplasm of Asparagus can be used in future improvement programs. The findings of current research on Asparagus germplasm can make a momentous contribution to initiatives of interbreeding, conservation, and improvement of Asparagus in future.

The global COVID-19 pandemic caused by SARS-CoV2 infected millions of people and resulted in more than 4 million deaths worldwide. Apart from vaccines and drugs, RNA silencing is a novel approach for treating COVID-19. In the present study, siRNAs were designed for the conserved regions targeting three structural genes, M, N, and S, from forty whole-genome sequences of SARS-CoV2 using four different software, RNAxs, siDirect, i-Score Designer, and OligoWalk. Only siRNAs which were predicted in common by all the four servers were considered for further shortlisting. A multistep filtering approach has been adopted in the present study for the final selection of siRNAs by the usage of different online tools, viz., siRNA scales, MaxExpect, DuplexFold, and SMEpred. All these web-based tools consider several important parameters for designing functional siRNAs, e.g., target-site accessibility, duplex stability, position-specific nucleotide preference, inhibitory score, thermodynamic parameters, GC content, and efficacy in cleaving the target. In addition, a few parameters like GC content and dG value of the entire siRNA were also considered for shortlisting of the siRNAs. Antisense strands were subjected to check for any off-target similarities using BLAST. Molecular docking was carried out to study the interactions of guide strands with AGO2 protein. A total of six functional siRNAs (two for each gene) have been finally selected for targeting M, N, and S genes of SARS-CoV2. The siRNAs have not shown any off-target effects, interacted with the domain(s) of AGO2 protein, and were efficacious in cleaving the target mRNA. However, the siRNAs designed in the present study need to be tested in vitro and in vivo in the future.

Nanomedicine has evolved as precision medicine in novel therapeutic approach of cancer management. The present study investigated the efficacy of biogenic gold nanoparticles synthesized using Argemone mexicana L. aqueous extract (AM-AuNPs) against the human colon cancer cell line, HCT-15. Biosynthesis of AM-AuNPs was determined by ultraviolet-visible spectroscopy and further characterized by transmission electron microscopy, X-ray diffraction, and Fourier transition infrared spectroscopy analysis. The cytotoxic activity of AM-AuNPs was assessed by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, whereas genotoxicity was evaluated by the DNA fragmentation assay. The expression of apoptosis regulatory genes such as p53 and caspase-3 was explored through semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and western blotting to evidence apoptotic cell death in HCT-15 cells. Biogenic AM-AuNPs inhibited cell proliferation in HCT-15 cell line with a half maximal inhibitory concentration (IC50) of 20.53 μg/mL at 24 h and 12.03 μg/mL at 48 h of exposure. The altered cell morphology and increased apoptosis due to AM-AuNPs were also evidenced through nuclear DNA fragmentation and upregulated expression of p53 and caspase-3 in HCT-15 cells. The AM-AuNPs may exert antiproliferative and genotoxic effects on HCT-15 cells by cell growth suppression and induction of apoptosis mediated by activation of p53 and caspase-3 genes.