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All Journal Livestock and Animal Research Livestock and Animal Research
Endang Tri Margawati
Research Center for Biotechnology, Indonesian Institute of Sciences
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Veterinary

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The Effects of Temperature and Culture Medium Composition on Protein Recombinant JSU-pET Expression Indriawati Indriawati; Slamet Diah Volkandari; Endang Tri Margawati
Sains Peternakan: Jurnal Penelitian Ilmu Peternakan Vol 17, No 2 (2019): Sains Peternakan
Publisher : Universitas Sebelas Maret (UNS)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (652.074 KB) | DOI: 10.20961/sainspet.v17i2.30374

Abstract

There are several factors influencing the protein expression. The study was aimed to increase the yield of protein recombinant JSU-pET by applying combination of temperature and medium culture composition. This research was composed in a 2x2 factorial design with two level temperatures of 37oC and 25oC and 2 medium compositions of LB and mLB (modified). Escherichia coli bearing JSU-pET plasmid were grown in two applied media and incubated in two applied temperatures. The cells were harvested by centrifugation the pellets were then lysed. The lysed cells were centrifuged, the supernatant then purified by Ni-NTA Resin Agarose. The purified JSU protein recombinant was characterized and quantified by spectrophotometer. The Result showed that the JSU protein recombinant was successfully expressed on the right size (37kDa) in all treatments even though the JSU-pET showed sharper band in m LB at 37oC. Statistically the protein yield was not significant different (P>0.05), however culture temperature of 25oC showed higher yield (0.618±0.095 vs 0.704±0.094) than cultured at 37oC (0.598±0.137 vs 0.553±0.041), respectively either cultured in LB or mLB. This finding suggests that a lower culture temperature could increase protein yield both either in LB or modified LB.
Detection of the CD18 gene mutation as a marker of BLAD genetic disorder of Holstein-Friesian cattle in West Java Mukh Fajar Nasrulloh; Ari Sulistyo Wulandari; Indriawati Indriawati; Endang Tri Margawati; Slamet Diah Volkandari
Livestock and Animal Research Vol 18, No 2 (2020): Livestock and Animal Research
Publisher : Universitas Sebelas Maret

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (422.154 KB) | DOI: 10.20961/lar.v18i2.42933

Abstract

Objective: Bovine Leukocyte Adhesion Deficiency (BLAD) is a genetic disorder in Holstein-Friesian (HF) cattle that have a significant economic impact on dairy cattle breeding. Mutation c.383A>G in Cluster of Differentiation Molecule 18 (CD18) gene was known as related to BLAD. This study aimed to detect the CD18 gene mutation that causes BLAD of HF cattle in the West Java Province.Methods: The genomic DNA was extracted from 88 blood samples of HF cattle from Cibungbulang-Bogor (n=34), Ciampea-Bogor (n=31), and Sukabumi (n=23) by the GB300 DNA extraction kit, GeneaidTM. The CD18 gene mutation was detected by PCR-RFLP analysis using the TaqI restriction endonuclease.Results: All samples in this study produced two fragments DNA, i.e., 359 bp and 260 bp with the monomorphic homozygote genotype (BB). HF cattle with BB genotype indicated as normal cattle and did not carry BLAD.Conclusions: All samples in the three populations of West Java were free of BLAD genetic disorder. Identification of BLAD on HF cattle in other places, including bulls in the Center of Semen Production in Indonesia was needed to prevent this genetic disorder.
Co-Authors Ari Sulistyo Wulandari Indriawati Indriawati Mukh Fajar Nasrulloh Slamet Diah Volkandari