Transposon is well-known as mobile element found abundant both in prokaryote and eukaryote genomes. In bacteria, transposon (famous name of a transposable DNA) could jump from chromosome to plasmid and its contrary. One type of transposons in bacteria known as insertion sequence (IS), it does not contain any additional genes except a gene encoding transposase, an enzyme that correlated to transponsition activities. The finding of transposon insertion unfortunately found during cloning of a fragment derived from deep-sea metagenomic DNA in this research. In the initial, this research was aimed to clone and characterize the á-amylase encoded gene derived from metagenomic DNA isolated from deep-sea water and sediment of Kawio Island, North Sulawesi. Metagenomic DNA has been isolated from deep-sea water and sediment and by using Whole Genome Amplification (WGA) technique, the DNA it could be increased in quantities to 146,31 ng for each 1 ng of metagenomic DNA. A fragment of ~1000 bp in length was obtained by using touchdown PCR method. The presence of a transposon in this DNA fragment is proposed as a hypothesis for losing ~700 bp leaving just 310 bp cloned sequence. Analysis of sequencing result showed a highest similarity between this 310 bp partial fragment with a replication protein (Rep) encoded gene from Pseudomonas putida (Query Coverage: 88%; Max. Identity: 80%, Positive: 86%) and this protein is known to be involved in plasmid replication where transposase encoding genes known usually presence together with this gene (Rep gene) in a bacterial plasmid.
One of the important enzymes in cellulase complex is β-glucosidase. In this research, adding signal peptide of inulinase gene from Kluyveromyces marxianus, cloning, and expressing of bglp15.2 gene in S. cerevisiae BY4741 had been done. Gene of bglp15.2 encoding β-glucosidase has 90% identity to nucleotide sequence of Shewanella frigidimarina NCIMB 400 bacteria. Adding nucleotide sequence of signal peptide was aimed to secrete β-glucosidase and had been done with PCR (Polymerase Chain Reaction) method. The addition of nucleotide sequence of signal peptide in bglp15.2 gene had been done succesfully that indicated from nucleotide sequencing result and the increment of amplicon band size in electroferogram of the last addition PCR step. The bglp15.2 and bglp15.2INU gene (the bglp15.2 gene that has signal peptide nucleotide sequence) were cloned in Escherichia coli DH5α using pGEM-T-Easy vector and pBEVY-GL shuttle vector. The pBEVY-GL shuttle vector was used for transforming S. cerevisiae BY4741 with bglp15.2 and bglp15.2INU. The recombinant S. cerevisiae BY4741 carrying bglp15.2INU gene and growing in 48 hours had extracellularly β-glucosidase enzyme activity of 0,0178 U/ml and the intracellularly activity was 0,0181 U/ml. The β-glucosidase enzyme without signal peptide was not secreted. With K. marxianus inulinase signal peptide, about 50% Bglp15.2INU protein could be secreted. The protein molecular weight of secreted Bglp15.2INU was 44 kDa in SDS-PAGE result.
Coronavirus disease 2019 (COVID-19) first appeared in China in December 2019 and was declared a pandemic by the World Health Organization. COVID-19 is caused by Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2), a new virus previously unknown to humans. Here we look at what is known about this virus, the main method for detecting the presence of this virus in a person who is used as a golden standard, and the problems that could arise in this detection method. Understanding the biology of the virus and the strengths and weaknesses of the detection method are important for patient management and for overcoming the pandemic.
The face is an important visual stimulus in daily life and each face identifies a particular person. The bone structure of the skull along with various soft tissues and coloration influence perception of the face. Facial averageness, and bilateral symmetry are the two most commonly used criterion of facial attractiveness, yet, both may be perceived differently based on hormonal status of the person observed. Facial perceptions may also differ according to cultural norms. In this research, we examined variations in face-shape among Sundanese male and female adults aged 18 to 40. We applied geometric-morphometric methods to analyze the landmark-based morphological variations in the frontal and lateral views of subjects’ faces. We identified five types of female frontal face views and four of male. We also identified five types each of female and male lateral face views. The trichion, gonion and gnathion were three most variable landmarks among the face views in our study, and highly determined the shape of the individuals’ faces. Multiple face type variation may refer to many categories of attractive faces since there is no exactly perfect category in the assessment of facial attractiveness by the viewers. Therefore, we believe that the configuration of facial features cannot constitute the sole visual criterion of facial attractiveness.
Banana ripening is a complex molecular process that produces visible changes in the texture, aroma, taste and nutritional content. Ripening is controlled by genetic code, metabolic pathway and associated microbiome. We reported the microbial community structure during banana ripening with alcohol treatment to discover endophytic and epiphytic microbes. We observed the pulp and peel from the first and seventh days of Cavendish (Musa acuminata cv. Cavendish) from mature green fruit and treated with 70% alcohol or distilled water sum up to eight samples and applied the 16S rRNA Illumina sequencing from V3–V4 gene region. After quality check 144,368 sequences were obtained in the dataset comprising a total read length of 1,237,805 base pairs. A sum of 199 genera were successfully isolated, with genera Alcaligenes was the most dominant genera at 56.65% and followed by more than 1% were genera Acinetobacter, Enhydrobacter, Pseudomonas, Stenotrophomas, Thermus, and Aerococcus using mothur pipelines. The highest diversity sample with 101 unique genera was belongs to distilled water treated raw bananas peel (NN1K) and the lowest diversity at 38 was belongs to distilled water treated ripe bananas pulp (NN7D). The metagenome data are available at NCBI Sequence Read Archive (SRA) database and Biosample under accession number PRJNA590572. The data contribute to discover different bacterial communities during post-harvest treatment. Keywords: Amplicon sequencing, Musa acuminata AAA group, Endophytic, Epiphytic, Alcaligenes