Penelitian molekuler untuk menemukan gen pengkode resistensi Multidrug Resistance Prtotein A (MRPA) T. evansi dan perbanyakan gen secara Polimeration Chain Reaction (PCR) masih sedikit dilakukan dan sangat penting untuk dipahami oleh mahasiswa calon guru biologi. Kajian ini bertujuan untuk menganalisis proses desain dan optimasi primer untuk gen target MRPA T. evansi yang dapat digunakan sebagai sumber belajar mahasiswa pendidikan biologi. Penelitian ini merupakan penelitian deskriptif mengenai tahapan mendesain primer secara online, optimasi primer secara laboratorium serta kajian mengenai pentingnya penerapan hasil studi ini dalam pembelajaran. Hasil penelitian menunjukkan terdapat tiga desain primer yang memenuhi syarat, selanjutnya dari tiga primer tersebut hasil optimasi di laboratorium menunjukkan hanya terdapat dua primer yang menunjukkan hasil yang baik dan dapat digunakan untuk penelitian amplifikasi gen MRPA T. evansi, yaitu primer pertama (F1’, R1’) dan primer kedua (F2’, R2’). Hasil kajian desain dan optimasi primer ini menunjukkan bahwa mahasiswa pendidikan biologi sangatlah penting untuk memahami konsep terkait dengan pekerjaan molekuler seperti mendesain dan optimasi primer, dikarenakan mereka memiliki tuntutan untuk menjadi seorang calon pendidik atau sebagai calon peneliti dimasa depan.Design and Optimization of Trypanosoma evansi MRPA Primer Coding Genes and Application to Molecular Biology LearningAbstractMolecular research to find Multidrug Resistance Prtotein A (MRPA) resistance coding genes and gene propagation by Polimeration Chain Reaction (PCR) is still little done and is very important to be understood by prospective biology teacher students. This study aims to analyze the design and primary optimization process for the T. evansi MRPA target gene that can be used as a learning resource for biology education students. This research was a descriptive study to described the step of primer design and optimization due to the importance of this steps to be applied as learning source. The results showed that there were 3 primer designs that qualified, then after the optimizing step there were only two primers that showed a good result, the first primer (F1, R1) and second primer (F2', R2). The results of this study showed the importance of biology education students to understand the concepts related to molecular work because in the future they are not only become prospective educators, they also have demands as prospective researchers.
Background and Aim: Trypanosomiasis, also known as surra, is an infectious disease with a wide host spectrum. In Indonesia, this disease is caused by Trypanosoma evansi. Various trypanocidal drugs have been used to treat this pathogen and subsequent disease. Yet, the long-term trypanocidal administration generates drug-resistant T. evansi. Some have identified genetic alterations in T. evansi transporter protein-coding genes that may be responsible for drug resistance. The Multidrug Resistance Protein E (MRPE) gene is a likely candidate gene responsible for the individual resistance. To date, no research has focused on T. evansi MRPE (TevMRPE) in this context. Hence, this research aimed at analyzing and characterizing the TevMRPE gene and protein using a bioinformatics approach. Materials and Methods: T. evansi was isolated from buffalo suffering from surra in Ngawi Regency, Indonesia. Isolated T. evansi was inoculated and cultured in male mice. The T. evansi genome was isolated from mouse blood with a parasitemia degree as high as 105. A polymerase chain reaction procedure was conducted to amplify the putative MRPE coding gene. The amplicon was sequenced and analyzed using MEGA X, BLAST, and I-tasser softwares. Results: The putative TevMRPE coding gene showed sequence similarity as high as 99.79% against the MRPE gene from Trypanosoma brucei gambiense. The protein profile and characteristics depicted that the putative TevMRPE protein was related to a family of Adenosine Triphosphate-Binding Cassette (ABC) transporter proteins. This family of transporter proteins plays a crucial role in the resistance toward several medicines. Conclusion: The obtained gene sequence in this research was identified as the TevMRPE. This gene is homologous to the T. brucei gambiense MRPE gene and possesses ligand active sites for Adenylyl Imidodiphosphate. In addition, MRPE contains enzyme active sites similar to the cystic fibrosis transmembrane conductance regulator. These data suggest that ABC transport proteins, like MRPE, may be necessary to confer trypanocidal drug resistance in T. evansi.
Aedes aegypti mosquitoes was the primary vector for carrying dengue fever. Dengue vector control has been widely carried out to reduce the number of dengue cases in Indonesia, but resistance cases have occurred in several areas, so it was necessary to know the status of resistance to determine appropriate efforts to control the vector. The purpose of this study was to determine the resistance status of Aedes aegypti in Malang Regency to 0.05% Cypermethrin. This type of research was descriptive observational research. The research method used was the susceptibility method with WHO standards using impregnated paper with the insecticide 0.05% Cypermethrin. Aedes aegypti mosquitoes were obtained from ovitrap results in the form of eggs and larvae taken from rearing at the Chemical Laboratory of the University of Muhammadiyah Malang. The number of mosquitoes used was 400 mosquitoes of the Aedes aegypti species aged 3-5 days. Mosquitoes were contacted with impregnated paper 0.05% Cypermethrin for one hour and held for 24 hours. Based on WHO standards, the criteria for susceptibility were determined as follows: said to be vulnerable if mortality was >98%, said to be tolerant if mortality was between 80-98%, and said to be resistant if mortality was <80%. The results showed that Aedes aegypti in Malang Regency was tolerant to 0.05% Cypermethrin with death rate each district; Turen 90%, Kepanjen 90%, Karangploso 96%, and Dau 100%, with percentage of 94%. This research showed that Cypermethrin with a concentration of 0.05% was still effective in reducing the development of Aedes aegypti, but the rotation of use must still be considered to reduce the number of dengue cases.
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