Dwi Listyorini

Ciprofloxacin effect on the Fgf10 gene expression in chicken embryo (Gallus gallus domesticus) limb development

Ciprofloxacin is a 331.4 Dalton fluoroquinolone antibiotic with teratogenic potency due to its capability to penetrate the placenta. The previous study reported that Ciprofloxacin affected the body axis skeletal structure and autopodium in mice. This study aimed to determine the effects of Ciprofloxacin on the expression of Fgf10 which regulates the development of chicken embryos (Gallus gallus domesticus) limbs. The administration of 10 mg · kg−1 BW (body weight) Ciprofloxacin had been done over five embryos of experimental group during day second to fourth of incubation. The isolation of total RNA from both wings and legs was performed at day fifth. The quantitative expression level analysis of Fgf10 gene was done through qRT-PCR. Limbs skeletal structure of 11 d embryos stained in Alcian Blue and Alizarin Red were observed as a supporting data. The results showed that Ciprofloxacin did not significantly suppress Fgf10 expression nor further investigation is required to reveal the effect of Ciprofloxacin on the genetic mechanism of both bone morphogenesis and limb development.Ciprofloxacin is a 331.4 Dalton fluoroquinolone antibiotic with teratogenic potency due to its capability to penetrate the placenta. The previous study reported that Ciprofloxacin affected the body axis skeletal structure and autopodium in mice. This study aimed to determine the effects of Ciprofloxacin on the expression of Fgf10 which regulates the development of chicken embryos (Gallus gallus domesticus) limbs. The administration of 10 mg · kg−1 BW (body weight) Ciprofloxacin had been done over five embryos of experimental group during day second to fourth of incubation. The isolation of total RNA from both wings and legs was performed at day fifth. The quantitative expression level analysis of Fgf10 gene was done through qRT-PCR. Limbs skeletal structure of 11 d embryos stained in Alcian Blue and Alizarin Red were observed as a supporting data. The results showed that Ciprofloxacin did not significantly suppress Fgf10 expression nor further investigation is required to reveal the effect of Ciprofloxaci...

Expression of gene encoded homogentisate geranylgeranyl transferase involved in tocotrienol biosynthesis in Indonesian local rice (Oryza sativa Linnaeus)

Banyuwangi has various kinds of local rice varieties, including Hitam Melik, Merah Harum, and Genjah Harum. These rice predicted to contain an antioxidant, such as vitamin E or tocochromanol (tocotrienol and tocopherol). Tocotrienol biosynthesis catalyzed by homogentisate geranylgeranyl transferase (HGGT) enzyme. The objective of this study was to determine the expression of gene encoded HGGT. Homogentisate geranylgeranyl transferase (HGGT) gene expression was studied by Quantitative Polymerase Chain Reaction (qPCR) and analyzed using comparative C(T) method or 2(ΔΔCt) method. HGGT gene expression results in Hitam Melik is higher than in Genjah Harum and Merah Harum. Total tocotrienol contents analyzed using High Performance Liquid Chromatography (HPLC) methods. There are three types of tocotrienol isoforms: α-tocotrienol; γ-tocotrienol; and δ-tocotrienol. Each of the rice varieties showed the different value of tocotrienol, specifically: Hitam Melik (823.54 µg · mL−1); Genjah Harum (692.28 µg · mL−1); and Merah Harum (586.84 µg · mL−1). Total tocotrienol measured by HPLC is in accordance with the gene expression results.Banyuwangi has various kinds of local rice varieties, including Hitam Melik, Merah Harum, and Genjah Harum. These rice predicted to contain an antioxidant, such as vitamin E or tocochromanol (tocotrienol and tocopherol). Tocotrienol biosynthesis catalyzed by homogentisate geranylgeranyl transferase (HGGT) enzyme. The objective of this study was to determine the expression of gene encoded HGGT. Homogentisate geranylgeranyl transferase (HGGT) gene expression was studied by Quantitative Polymerase Chain Reaction (qPCR) and analyzed using comparative C(T) method or 2(ΔΔCt) method. HGGT gene expression results in Hitam Melik is higher than in Genjah Harum and Merah Harum. Total tocotrienol contents analyzed using High Performance Liquid Chromatography (HPLC) methods. There are three types of tocotrienol isoforms: α-tocotrienol; γ-tocotrienol; and δ-tocotrienol. Each of the rice varieties showed the different value of tocotrienol, specifically: Hitam Melik (823.54 µg · mL−1); Genjah Harum (692.28 µg · mL−1); an...

Isolation of Flower Development Regulator Gene SEPALLATA 1 in Phalaenopsis amabilis (L.) Blume

Phalaenopsis amabilis (L.) Blume is an indigenous orchid species in Indonesia. This orchid has a white large flower. The large flower is caused by the existence of gene that has an important role in flower development. One of the genes is SEPALLATA 1. This gene is a member of superfamily MADS-Box gene. SEPALLATA 1 gene is a marker of primordial flower organ. This study aimed to isolate SEPALLATA1 gene from Phalaenopsis amabilis (L.) Blume by PCR using forward primer 5’-GCT-GGA-GCG-GAT-CGA-GAA-CA-3’and reverse primer 5’-TCA-TGC-AAG-CCA-ACC-AGG-TG-3’ . This study successfully amplified 691 bp lengths of SEPPALATA1 fragment, lacking 20 bp upstream which consist its start codon. Keywords: Flower development regulation, Phalaenopsis amabilis (L.) Blume, SEPALLATA 1 gene.

The Phylogenetic Study of the White-Bellied Sea Eagle [Haliaeetus leucogaster (Gmelin, 1788)] Based on DNA Barcoding Cytochrome-c Oxidase Subunit I (COI)

Even though not yet considered as endangered, White-bellied Sea Eagle’s global population is decreasing due to illegal hunting, bird trading, and deforestation. So far, there hasn’t been any report regarding the phylogenetic study of the White-bellied Sea Eagle inhabiting the coastal regions of Java. Moreover, there hasn’t been any report on the genetic data, especially COI gene, of the White-bellied Sea Eagle living in coastal area of Java. Thus, in this research, two individuals of Heliaeetus leucogaster ( Gmelin , 1788); are compared based on its COI gene sequence to the member of genus Haliaeetu s to determine their position in the phylogenetic tree of genus Haliaeetus. COI gene amplification is performed using Forward primer BirdF1 5’- TTC TCC AAC CAC AAA GAC ATT GGC AC-3’ and Reverse primer BirdR2 5’ ACT ACA TGT GAG ATG ATT CCG AAT-3’. The phylogenetic analysis using MEGA6 with Maximum Likelihood method shows that Haliaeetus leucogaster in this study is related to Haliaeetus leucocephalus (Linnaeus, 1766), Haliaeetus pelagicus (Pallas, 1811), and Haliaeetus albicilla (Linnaeus, 1758). Keywords: phylogenetic study; Heliaeetus leucogaster ( Gmelin , 1788); DNA barcoding, C ytochrome-c O xidase S ubunit I (COI).

Pun1 Gene Isolation from Capsicum frutescens L. cv Cakra Hijau

Capsicum frutescens L. is one of chili peppers with high pungency. Capsicum frutescens has several cultivars, one of those is C. frutescens cv. Cakra Hijau. This cultivars is known resistance to pests and diseases as well. Pungency is due to the accumulation of capsaicinoids. Pun1 is an important gene responsible for pungency. The full-leght genomic sequence of Pun1 is 1897 bp, containing two exons of 738 bp and 590 bpand one intron of 348 bp in between. This study was aimed to isolate Pun1 gene that free from intron. mRNA was isolated with TriReagentâ furthermore RT-PCR method used Qiagent One-Step RT-PCR and two pairs of primer : F1/R1 (F15’-ATG-GCT-TTT-GCA-TTA-CCA-TCA-3’ / R15’- CTT-AGC-TCG-AAG-TGC-ATC-TA-3’) and F2/R2 (F25’-GAA-GGT-GGC-AGA-AGA-ATC-AG-3’/R25’-TTA-GGC-AAT-GAA-CTC-AAG-GA-3’). The result of this study are isolated 738 bp exon-1 and 590 bp exon 2 of Pun1 gene. Keywords: Capsaicin; Capsicum frutescens L. cv. Cakra Hijau; exon; Pun1 gene

Haplotype Diversity of COI Gene of Hylarana chalconota Species Found at State University of Malang

Hylarana chalconota is a cryptic species of frog endemic to Java Island [1]. This species is small with long legs, and brown skin. The Snout-Vent Length (SVL) ranges between 30-40 mm for male and 45-65 mm for female. [4] Reports the existence of this species in State University of Malang, which was not found in 1995 [5]. Sampel #1 displays spots in its skin, which does not exist in sample #2. To reveal the haplotype diversity of COI gene in this species, we analyzed Cytochrome-c oxidase subunit-1 (COI) sequences of both samples. Using a pair of primers according to [6] both samples had 604 bp and 574 bp fragment length, respectively. These fragments showed polymorphism; with mutation position in sites 104, 105, and 124. Based on this result, we suggest that the two samples share a different haplotypes, proposed as UM1 and UM2.