@article {2099, title = {Optimization of Osteopontin Recombinant Protein as a Candidate Supplementation for Semen Preservation}, journal = {Pharmacognosy Journal}, volume = {15}, year = {2023}, month = {August 2023}, pages = {668-673}, type = {Research Article}, chapter = {668}, abstract = {

The recombinant protein of heterologous proteins in Escherichia coli strains K12 has various and different systems tested and demands a detailed insight into the multiple factors affecting the encoded protein. One of the crucial factors is the acceptable quality of the DNA copies inserted inside the bacteria. Firstly, the amplification procedure needed to be performed well; thus, designing the primer and selecting the optimum annealing temperature are the focus indicators in this study. This study obtained a reference gene from the NCBI data bank with Reference Sequence: NM_174187.2. Two types of primers (SPP1FSPP1R and OPN1F - OPN1R) with different targeted bands were designed and selected after being reconstructed using the software. Online software such as addgene.org is also used to identify the right restriction site. The annealing temperature distinguished the PCR system used to amplify each primer. The result of this study revealed the best annealing at 65{\textordmasculine}C successfully amplified 820 bp of the targeted band. The phenomenon not following the theory of blue-white screening is the empty plasmid control, where not a single colony grows on the media. Competent cells inserted with empty plasmids should still be able to expand on LB-Amp agar media because the presence of these plasmids is capable of providing resistance to antibiotics (in this case, ampicillin). This discrepancy is thought to have been caused by the improper insertion of the empty plasmid so that the plasmid did not enter the competent cell.

}, keywords = {DNA clone, Osteopontin, PCR amplification., Primer}, doi = {10.5530/pj.2023.15.135}, author = {Tatik Hernawati and Tita Damayanti Lestari and Suzanita Utama and Rimayanti} } @article {1885, title = {Effect of Vitamin C and E Supplementation and Combination of Both in Egg Yolk Tris Diluter on the Quality of Sapera Goat Spermatozoa in the 5 {\textdegree}C Cooling Process}, journal = {Pharmacognosy Journal}, volume = {14}, year = {2022}, month = {October 2022}, pages = {661-665}, type = {Research Article}, chapter = {661}, abstract = {

This study was aimed to determine the effect of vitamin C, E supplementation and both combination in the egg yolk tris diluent on the quality of Sapera goat spermatozoa in 5oC cooling process. The samples were fresh semen collected from Sapera goat. It was divided into four different treatment groups. The control treatment or P0 contained semen sample + egg yolk tris diluent, P1 contained semen sample + egg yolk tris diluent + vitamin C 0,9 mg/ml, P2 contained semen sample + egg yolk tris diluent + vitamin E 1 mg/ml, P3 contained semen sample + egg yolk tris diluent + vitamin C 0,9 mg/ml + vitamin E 1 mg/ ml. The least decrease of spermatozoa motility, viability and membrane integrity was found in P3, which was significantly different to P0 (p\<0,05). However, P1 and P2 did not show significant difference in the results of motility and viability evaluation (p\>0,05). It can be concluded that the combination of vitamin C and E supplementation can be an optimal antioxidants supplement in egg yolk tris diluent on the quality of Sapera goat spermatozoa in 5oC cooling process.

}, keywords = {5oC Cooling Process, Quality of Spermatozoa, Sapera Goat, Vitamin C, Vitamin E}, doi = {10.5530/pj.2022.14.150}, author = {Amung Logam Saputro and Uliy Ba{\textquoteright}sin Syadid and Rimayanti and Suherni Susilowati and Ragil Angga Prastiya and Bodhi Agustono and Fauzan Mumtazi and Marifatunnisa{\textquoteright}romadhona and Anastasya and Muhammad Riesta Farhan} }