Open Access Original Research Article

Diversity Analysis of Sweet Potato (Ipomoea batatas [L.] lam) Accessions from North Central Nigeria Using Morphological and Simple Sequence Repeats Markers

U. J. Alfred, C. C. Iheukumere, C. U. Aguoru, O. J. Olasan, U. M. Sesugh

Asian Journal of Biotechnology and Genetic Engineering, Page 1-15

Aims: Genetic diversity analysis was carried out with the aim of assessing the genetic similarities and variability that existed among sweet potato accessions that are grown in North Central Nigeria using Morphological and Simple Sequence Repeats makers.

Study Design: The field experiment was laid out in a Randomized Complete Block design (RCB) with 5 replications. Morphological characterization was done in the field while Molecular characterization was carried out in the Molecular laboratory.

Place and Duration of Study: Field experiment was carried out at the Teaching and Research farm of the Federal University of Agriculture Makurdi while the laboratory experiment was done at the Molecular Biology laboratory of the Federal University of Agriculture Makurdi. The experiment was carried out between May and August 2018.

Methodology: A total of 20 potato accessions collected from six states (Benue, Kogi, Nasarawa, Niger, Plateau and Abuja) in north central Nigeria were planted in the field for morphological characterization and Observations were made on 21 morphological characters at 90 days after planting (DAP). Genomic DNA for molecular characterization was extracted from young leaves (20 DAP) located at the tip of the main vine of the sweet potato plant using DNA Zol extraction protocol. The extracted DNA was amplified using five SSR primers via Polymerase Chain Reaction in a thermocycler. The amplified DNA was then subjected to 5% Agarose gel electrophoresis and the products were viewed under U-V light. The bands formed as a result of amplification were scored in a binary pattern for analysis.

Results: ANOVA of the of morphological characters revealed that there were significant variation for 18 out of the 21 morphological characters studied among the sweet potato accessions, with the first 4 Principal Components accounting for 72.1% of the total variations among the accessions. The 18 characters were thus useful as morphological markers for diversity analysis and based on them, cluster analysis grouped the sweet potato accessions into 4 clusters based on their average linkages and the Euclidean test. Four pairs of duplicates (NC 7 and NC 15, NC 8 and NC 16, NC 17 and NC 19, NC 18 and NC 20) were identified to be similar accessions based on the morphological characterization. For molecular characterization, Polymorphic Information Content (PIC) for the DNA bands formed showed the usefulness of the primers used in revealing genetic diversity among the accessions with primer 1 (IB02) and 4 (IBS 199) showing 31.818% polymorphism in 1 (IB02) and 4 (IBS 199). For cluster analysis, three distinct clusters were observed with all the accessions in cluster I and II which were approximately 30% similar while accession NC 10 which stood alone in cluster III shared no similarity with any of the other accessions and could possibly be a hybrid. The cluster analysis also revealed a total of 4 sets of duplicates thereby further reducing the total number of accessions to 6 which indicated that a very low diversity existed among the accessions.

Conclusion: This study established that a lot of duplicates existed among the sweet potato accessions indicating that there is a very low level of sweet potato genetic diversity in the North Central region of Nigeria.

Open Access Original Research Article

Investigation of SKOR2 Gene Expression in Patients with Lung Cancer Using Real-time PCR

Mohammadreza Ghanbari, Seyed Mohammad Amin Ahmadpanah, Fahimeh Nemati

Asian Journal of Biotechnology and Genetic Engineering, Page 1-7

Background and Aims: Lung cancer includes small cell lung cancer and non-small cell lung cancer (NSCLCs) types. NSCLCs are investigated for various mutations, which is available can be treated with targeted new molecular therapies. The aim of this study was to evaluate the expression of SKOR2 gene, disease severity, and evaluation of NSCLC and its subtypes of cancer patients among patients with lung cancer in Tehran hospitals using Real-Time PCR.

Materials and Methods: A total of 35 clinical samples were collected from patients with NSCLC-derived lung cancer from three hospitals in Tehran. The range of patients varied from 37 to 80 years. The disease grade ranges in the patients in this study were varied and 22 different grades were observed. In order to evaluate the SKOR2 gene after extraction of RNA and cDNA synthesis, the gene expression was evaluated using Real-Time PCR.

Results: Of the 22 observed grades, the highest grade IIIa grade was observed in 6 patients (17.1%). 74% of adenocarcinoma cases were in T-categories of lung cancer and 25% of patients were in grade IIIa. Patients with T3 stage included 4 samples, 2 of which were adenocarcinoma and 2 were SCC and their age ranged from 55 to 62 years old. Three patients were in the T1 category, and 100% of them had adenocarcinoma. The results showed that expression of SKOR2 5.47 fold in lung cancer patients with NSCLC was more than normal patients.

Conclusion: According to the results, expression of the SKOR2 gene in the tissues of individuals with lung cancer was increased compared to normal individuals. The expression of this gene in patient suggests the possibility of involvement of this agent in the progression of the disease. According to the results, it is recommended to use an evaluation of the expression of other biomarkers in lung cancer to help with this and to increase the accuracy of screening tests for lung cancer.

Open Access Original Research Article

Study of ZNF804A and DISC1 Genes in Iranian Patients with Schizophrenia

Azizi Faryal, Azizi Parisa, Nemati Fahimeh

Asian Journal of Biotechnology and Genetic Engineering, Page 1-6

Background: Schizophrenia is a complex and chronic psychological disorder characterized by a set of symptoms including illusions, speech and behavioral disorders, and cognitive impairment. Schizophrenia is a complex multiple disorder, and the biggest risk factor for it is a positive family history. The aim of this study was to investigate the relationship between polymorphism of ZNF804A and DISC1 genes in patients with schizophrenia in Iran.

Materials and Methods: In this case-control study, 50 patients with schizophrenia and 50 healthy controls were evaluated. The PCR-RFLP method was used to evaluate single nucleotide polymorphism in both groups of patients and control. For enzymatic digestion of PCR products, rs1344706 and rs6675281 were amplified and digested using MboI and BseLI enzymes at 37 ° C for 16 hours, respectively.

Results: The frequency of TT, GT and GG genotypes for ZNF804A gene in rs1344706 was 26%, 52%, and 22%, respectively, and in healthy subjects 46%, 42%, and 8%, respectively. In the DISC1 gene, the frequency of TT, CT and CC genotypes in the rs6675281 region was 2%, 14%, and 84%, respectively, and 2%, 14%, and 80%, respectively, in healthy subjects or controls, respectively.

Conclusion: Frequency of homozygous GG and heterozygote GT genotypes was 8% and 14% higher than healthy subjects, but the frequency of homozygous TT in healthy subjects was 22% higher than those with schizophrenia for ZNF804A gene in rs1344706 region. However, in case of DISC1 gene, the frequency of TT, CT and CC genotypes in the rs6675281 region was very similar in healthy and healthy subjects, and there was no significant difference between homozygous and heterozygous genotypes. Therefore, the result of our study can be a way to providing suitable information about the disease in order to prepare patients and family and to program adjusted treatment to prevent major injuries.

Open Access Original Research Article

Antidiarrhoeagenic Potentials of Synergistic Activities of Water Extracts of Aloe vera and Hyptis suaveolens against Giardia lamblia and Salmonella Species Infections among Children 0-5 Years in Bauchi State, Nigeria

Omammegbe Abdulrahman, Adamu B. Samaila, S. M. Panda, Abdullahi Aliyu, M. R. Sahal

Asian Journal of Biotechnology and Genetic Engineering, Page 1-7

Background: The epidemiological significance of Salmonella species and Giardia lamblia co-infections of childhood diarrhoea differs substantially between regions depending on socio-economic conditions in the prevalence of different categories of diarrhoeagenic disease associated with diarrhoea among under 5 populations, medicinal plants extracts has increasingly been advocated in productions of antimicrobial agents as resistance with synthetic drugs increases.

Methodology: The phytochemicals constituents obtained from Aloe vera and Hyptis suaveolens were investigated to evaluate their medicinal potentials, the screening revealed the presence of saponins, tannins, alkaloids, flavonoids, terpenoids, alkaloids, phenols. The study was conducted between June, 2018 and February, 2019 in an attempt to ascertain the efficacies of Aloe vera and Hyptis suaveolens individually and in combined against diarrhoea infections caused by Giardia lamblia and Salmonella species in under five children in Bauchi State, Nigeria. Anti-giardial activity was carried out in an in-vitro susceptibility assays method on Giardia lamblia and antibacterial activity was carried out by Kirby-Bauer method on Salmonella species. Parasites mortality was determined by counting in hemocytometer under a light microscope and zone of inhibitions produced against the bacteria were determined and subjected to descriptive statistics and inferential statistical analysis to determine their significance at 5% level using SPSS version 23. Means were separated using DMRT (P≤0.05).

Results: The results obtained showed that water extracts of Aloe vera and Hyptis suaveolens plants singly and in combinations had inhibitory effects on Giardia lamblia and Salmonella species. Shows that Aloe vera extracts on Salmonella species exhibited good zone of inhibitions 0.302±18.00, Hyptis suaveolens extracts 0.315±19.67 and the combined extracts 0.413±30.00. On Giardia lamblia, shows better activities with Aloe vera extracts 0.002±0.505, Hyptis suaveolens extracts 0.002±0.478 and the combined extracts 0.002±0.643. In all the cases, plants, concentrations and time were determinants factors for the anti-giardial and antibacterial activity.

Conclusion: Hence, Hyptis suaveolens extracts have better activities than extracts of Aloe vera while the combined extracts shows more better activities of antibacterial, Aloe vera have better activities than Hyptis suaveolens extracts while the combined extracts shows more better activities of antigiardial. Therefore, these plants possess antimicrobial potentials.

Open Access Review Article

CRISPR-Cas9: A Detail on Specific Tool for Gene Editing or Targeting

Lokendra Singh

Asian Journal of Biotechnology and Genetic Engineering, Page 1-7

Objective: The applicability in reverse genetics studies, animal model experiments and also in curing diseases caused by viruses like HIV makes the CRISPR-Cas9 system a highly desirable technique with the ability to achieve quick plausible outcomes. The bacterial cells chop up the viral genome. And its search of these pieces of spacer DNA. The spacer DNA is nothing but pieces of the different viral genome. That had infected the cell previously. So each time of bacteriophage infects the cell take it pieces and search of spacer DNA. The interference is between the specific Cas protein and RNA Sequence has going to be merged together. The Crispr system would be somehow like adopted immune system in human. it would kind of memory to prevent this same bacteriophage for infecting the cell other time.

Purpose (Hypothesis): This article main purpose to describe the Genome editing enabled by the development of tools to make precise, targeted changes to the genome of living cells.

Methods (Technique): CRISPR-Cas9.

Conclusion: The CRISPR-Cas9 is the key to genetic revaluation era. This tool is very useful for studying the genetic code. at finally in this review both in modelling and the CRISPR/Cas9 system have emerged as a feasible tool to achieve what has seemed impossible for decades.