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All Journal BERITA BIOLOGI Animal Production : Indonesian Journal of Animal Production
WT Artama, WT
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Veterinary

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Expression and Isolation of Recombinant Microneme 3 (MIC3) Protein of Toxoplasma gondii Local Isolate on Eschericia coli (BL21) Indrasanti, D; Haryanto, A; Artama, WT
ANIMAL PRODUCTION Vol 13, No 2 (2011): May
Publisher : Universitas Jenderal Soedirman, Faculty of Animal Science, Purwokerto-Indonesia

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Abstract. Toxoplasmosis is a disease that infects all warm-blooded animals, including livestocks and humans caused by Toxoplasma gondii parasites. There are major drugs used for the therapy, though they have some effects to the patients, such as allergy, toxic and teratogenic for fetus. In addition, toxoplasmosis treatment is only effective for tachyzoites T. gondii in acute infection, while tissue cysts cannot be eradicated in chronic toxoplasmosis Tissue cysts of T. gondii contained in meat that are consumed by humans and meat-derived products may be important sources of infection for humans. Microneme protein (MIC) is one of proteins that belongs to excretory-secretory antigens (ESAs) of Toxoplasma gondii. Microneme 3 protein (MIC3) is the protein that plays an important role in the invasion process during cell infection as a mediator attachment parasite to the host cell. Recombinant MIC3 protein has been already used for the detection of toxoplasmosis and it could induce humoral and cellular immune response in experimental animals. The aim of this research was to express MIC3 recombinant protein of T. gondii from local isolate that was cloned into expression vector and transformed to E. coli BL21. In the future, recombinant protein MIC3 can be used for vaccine candidate and diagnostic tools for toxoplasmosis in animals and humans. Gene of MIC3 T. gondii local isolate (1.2 Kbp) was cloned into expression vector pET-32a(+) (5.9 Kbp) and transformed to Escherichia coli BL21. Protein from plasmid recombinant (7.1 Kbp) was expressed and performed by culturing recombinant bacteria into LB medium containing ampicillin and IPTG. Recombinant protein was isolated by sonication method and identified using SDS-PAGE. Finally, the recombinant protein was analyzed by immunoblotting using anti-ESAs polyclonal antibody. In conclusion, expression of the MIC3 gene with ~108 kDa has been successfully performed by cloning gene encoding for MIC3 protein of T. gondii local isolate that can be identified with polyclonal antibody anti-ESAs.Key Words: Toxoplasma gondii, expression, MIC3 protein
RFLP Marker Variation of Cytocrome b Gene and Genetic Relationship among Batur, Merino and Local Sheep Breeds Prayitno, Prayitno; Hartati, T; Pratiwi, R; Artama, WT
ANIMAL PRODUCTION Vol 13, No 3 (2011): September
Publisher : Universitas Jenderal Soedirman, Faculty of Animal Science, Purwokerto-Indonesia

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RFLP analysis of mitochondrial DNA cytochrome b gene was conducted to determine the diversity, status and close genetic relationships in a population of Batur sheep with the Merino and local sheep breeds (Garut, Thin Tail and Fat Tail). The research used genomic DNA of 27 samples of Batur, 15 Merino, 17 Garut, 15 Thin Tails and 15 Fat Tails sheep. The PCR process used two types of 25 nucleotides primers. The PCR products were checked by using 2% agarose gel electrophoresis. The PCR DNA fragment was digested by using Hae III at 37 oC and incubated for 10 hours. Similarities and differences of cytochrome b gene RFLP bands between individual samples of one and across populations, genetic distance, and close genetic relationship, were identified. The PCR process of the cytochrome b gene metochondrial DNA of the 45 samples of sheep yielded 359 bp band types. The digestion (cutting) of the PCR products of mitochondrial DNA cytochrome b gene by using Hae III resulted in RFLP band profiles of 128 up to 231 bp polymorphisms of cytochrome b gene. Although the Hae III restriction enzyme recognized only one restriction site, however, between samples of Batur, Merino, Garut, Thin Tail, and Fat Tails, there were monomorphism and polymorphism Hae III loci.Key Words:  RFLP, cytochrome b gene, genetic markers, genetic similarity, Batur sheeAnimal Production 13(3):156-165 (2011) 
Polymorphism of the Growth Hormone Gene and its Association with Growth Traits in Ongole Grade Crossed with Simmental Cattle Mu’in, MA; Astuti, M; Muladno, Muladno; Murti, TW; Artama, WT
ANIMAL PRODUCTION Vol 9, No 2 (2007): May
Publisher : Universitas Jenderal Soedirman, Faculty of Animal Science, Purwokerto-Indonesia

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The present study was conducted to detect polymorphism of growth hormone (GH) gene in Ongole grade crossed with Simmental bull (SIMPO) and its association with growth traits. Blood samples of 62 cattle were taken from population of SIMPO cattle were located in the sub-province region of Bantul, Special Region of Yogyakarta. A 211 bp fragment of GH gene spanning from the forth intron region (49 bp) to fifth of exon (162 bp) was amplified and digested with AluI restriction enzyme to identified polymorphism at this locus. The resulted indicated that two genotypes LL and LV were found at the GH gene in SIMPO population cattle. The frequencies of L and V alleles were 0.82 and 0.18, respectively. In SIMPO calves, the average birth weight, 2 months body weight and daily body weight gains of LV genotypes were tend to higher than that of LL genotypes. (Animal Production 9(2): 53-58 (2007) Key Words : Gene, growth hormone, polymorphism, cattle, SIMPO
Genetic Diversity of -Casein Gene of Friesian-Holstein Dairy Cattle using Restriction Fragment Length Polymorphism Technique Widayanti, R; Artama, WT; Subagyo, S; Winarso, D
ANIMAL PRODUCTION Vol 10, No 1 (2008): January
Publisher : Universitas Jenderal Soedirman, Faculty of Animal Science, Purwokerto-Indonesia

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The objective of the research was to evaluate the genetic varieties of k-Casein gene of Holstein Friesian (HF) in Koperasi Unit Desa (KUD) Karangploso, Dau, Ngantang and Pujon, Malang, East Java by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Analysis (PCR-RFLP) using Pst-l.  The research used Holstein Friesian dairy cows raised by farmers of KUD Karangploso, Dau, Ngantang and Pujon, Malang, East Java. The blood samples were collected from 29 cows. The research activities were carried out by collecting cow’s blood, DNA extraction, and DNA amplification with PCR. PCR products were digested with Pst-l enzyme restrictions, then allele genes of k-Casein were identified. The frequency of allele and genotype of k-Casein gene were calculated by Hardy-Weinberg. The results showed that cows raised by farmers of KUD Karangploso, Dau, Ngantang and Pujon, Malang-East Java had two alleles k-Casein polymorphism, i.e. allele A (0,74) and B (0,26); therefore, two genotypes existed including AA (0,55) and AB (0,38). It can be concluded that genetic diversity of k-Casein gene existed in Holstein-Friesian dairy cows raised by farmers of KUD Ngantang, Pujon, Dau and Karangploso, Malang, East Java. (Animal Production 10(1): 1-4 (2008) Key Words: Pst-1, k-casein gene, PCR-RFLP
Virgin Coconut Oil Increases the Productivity of Broiler Chicken Post Avian Influenza Vaccination Yuniwarti, EYW; Asmara, W; Artama, WT; Tabbu, CR
ANIMAL PRODUCTION Vol 14, No 3 (2012): September
Publisher : Universitas Jenderal Soedirman, Faculty of Animal Science, Purwokerto-Indonesia

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Abstract. Chicken productivity is not only determined by body weight increase and feed efficiency but also disease resistance. Avian influenza (AI) is still an endemic in Indonesia. Highly mutative characteristic of AI causes unsuccessful vaccination to preventing chicken mortality; therefore, feed modulation alternatives are sought to raise body weight and body immune as well. Virgin coconut oil (VCO) contains fatty acid potential as antimicrobe and antivirus; VCO intake is therefore expected to increase chicken body immune. This research aimed at feed modulation to increase broiler chicken productivity. Forty broiler chicken of one day old (DOC) were used and the research applied Completely Randomized Factorial Design in which factor one was two vaccine levels namely AI-vaccinated chickens and AI-unvaccinated chickens. Factor two used four levels of VCO: 0, 5, 10, 15 mL/kg feed. DOC chickens were divided into eight treatment groups and repeated in five experiment units. Feed and water were given ad libitum. The result demonstrated that in spite of heterophile increase in AI-vaccinated VCO-given chickens, heterophile/lymphocyte ratio and feed intake were not significantly different among all treatment groups. With the highest body weight found in AI-vaccinated chickens given 10ml/kg feed VCO, it could therefore be concluded that VCO intake of 10mL/kg feed could raise body weight.Key words: heterophile, heterophile/lymphocyte ratio, feed intake, body weightAbstrak. Produktivitas ayam tidak hanya ditentukan oleh kenaikan bobot badan dan efisiensi pakan, tetapi juga ketahanan terhadap penyakit. Avian influenza (AI) masih merupakan wabah endemis di Indonesia. Sifat AI yang mudah bermutasi menyebabkan vaksinasi tidak selalu berhasil untuk mencegah kematian ayam, sehingga dicari alternatif modulasi pakan untuk meningkatkan bobot badan dan kekebalan tubuh. Virgin coconut oil (VCO) mengandung potensi asam lemak sebagai antimikroba dan antivirus, sehingga asupan VCO diharapkan dapat meningkatkan kekebalan tubuh ayam. Penelitian ini bertujuan untuk modulasi pakan untuk meningkatkan produktivitas ayam broiler. Empat puluh ayam broiler umur satu hari  (DOC) digunakan dalam penelitian yang menerapkan Rancangan Acak Lengkap Faktorial dengan faktor pertama dua level vaksinasi  yaitu ayam divaksin AI dan tidak divaksin AI. Faktor kedua adalah empat level VCO: 0, 5, 10, 15 mL  kg pakan. DOC ayam dibagi menjadi delapan kelompok perlakuan dan diulang dalam lima unit percobaan. Pakan dan air minum diberikan ad libitum. Hasilnya menunjukkan bahwa meskipun adanya kenaikan heterofil pada ayam yang divaksin AI dan diberi VCO, rasio heterofil/limfosit dan konsumsi pakan tidak berbeda secara signifikan pada semua kelompok perlakuan. Dengan bobot badan tertinggi ditemukan pada ayam yang divaksin AI dan diberi VCO 10 ml/kg pakan, maka dapat disimpulkan bahwa asupan VCO 10 mL/kg pakan dapat meningkatkan bobot badan. Kata kunci: heterophil, rasio heterophil/limfosit, konsumsi pakan, bobot badan EYW Yuniwartiet al/Animal Production 14(3):192-198, September 2012
Genetic Relatedness Between Batur, Merino and Local Sheep Based on Random Amplyfied Polymorphism DNA Marker Prayitno, Prayitno; Hartatik, T; Pratiwi, R; Artama, WT
ANIMAL PRODUCTION Vol 13, No 1 (2011): January
Publisher : Universitas Jenderal Soedirman, Faculty of Animal Science, Purwokerto-Indonesia

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RAPD analysis to determine the diversity, status and genetic close relatioship Batur with Merino, Garut, Thin Tail and Fat Tail sheep genomic DNA was used in 27 Batur, 15 Merino, 17 Garut, 15   Thin Tails and  Fat Tails animals. The process of RAPD-PCR used five primers of 10 nucleotides. PCR results were electrophoresed with 2% agarose gel. Identified similarities and differences between individual RAPD bands one and cross-sample of the population, genetic distance and closeness of genetic relationship. The process 89 sample  sheep RAPD-PCR generated of 4189 band from 100 to 1500 bp which consisted of 91 type. Batur sheep samples produced bands at most (1666 tape) and the lowest Fat Tails (493 bands). The number monomorfism of bands most of the Batur (891 bands) and the lowest Garut (170 bands), but the polymorphism  bands highest  of the Batur (775 bands) and the lowest Fat Tails (262 bands). Between individuals within populations have similar genetic Merino highest and the lowest Thin Tail. Cross-population genetic similarity is highest individuals in the population Batur and Merino, while the lowest Merino and Thin Tail. The closest genetic distance was Batur with Merino population and the most apart distant Merino with Thin Tail or Merino and Fat Tails. Batur sheep population has the closest genetic relationship with the Merino and the most apart distant is with Fat Tail. (Animal Production 13(1):30-38 (2011)Key Words: RAPD, genetic markers, genetic similarity, sheep
ANALISIS IMUNOGENISITAS PROTEIN GRA1 DARI HASIL KLONING GEN GRA1 TAKIZOIT Toxoplasma gondii [Immunogenicity Analysis of GRA1 Protein Derived from Clone Bearing GRA1 Genes Collected from Toxoplasma gondii Tachyzoite] Subekti, Didik T; Artama, WT; Poerwanto, SH; Sulistyaningsih, E; Sari, Yulia
BERITA BIOLOGI Vol 11, No 1 (2012)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (269.729 KB) | DOI: 10.14203/beritabiologi.v11i1.482

Abstract

The study was aimed to analyze the immunogenicity of GRA1 protein derived from clone bearing GRA1 genes from local isolate of Toxoplasma gondii. Analysis of GRA1 protein translated from cDNA of GRA1 is very essential in prior to expressed the gene. Analysis of GRA1 protein derived from clone bearing GRA1 genes was performed using several bioinformatics software which are available as standalone or online software such as CLC Bio Workbench series, BioEdit, BESTORF, GENSCAN, FGENES, BepiPred 1.0, CTL Epitope Finder and SignalP. Translation coding sequences of GRA1 gene into GRA1 peptide sequences revealed 190 amino acids with molecular mass of GRA1 approximately 20.159 kD and isoelectric point at 4.43. GRA1 protein also identified several antigenic domains with six domains were known as epitopes for CD8+/cytotoxic lymphocyte and seven domain as epitopes for B lymphocyte. However, GRA1 protein was considered as good antigen but less immunogenic.
Co-Authors A Haryanto, A CR Tabbu, CR D Indrasanti, D D Winarso, D Didik T Subekti E Sulistyaningsih EYW Yuniwarti, EYW M Astuti, M MA Mu’in, MA Poerwanto, SH Prayitno Prayitno R Pratiwi, R R Widayanti, R S Subagyo, S T Hartati, T T Hartatik, T Tamaulina Br Sembiring TW Murti, TW W Asmara