Wesam Hasan Mohamed Mady1*, Bing Liu2, Dong Huang2, Abdel Satar Arafa1, Mohamed Khalifa Hassan1, Mona Mehrez Aly1, Pucheng Chen2, Yongping Jiang2* and Hualan Chen2
...ed into pCAGGS mammalian expression vector under the control of chicken β-actin promoter. The construct was transfected into 293T HEK (Human Embryonic Kidney) cell line for in vitro expression of the recombinant plasmid DNA. The confirmation of H5 protein expression was performed using SDS-PAGE followed by Western blotting and immunofluorescence assays. The humoral immune response was evaluated by intramuscular immunization of specific pathogen free (SPF)...
Tasawar Sultana*, Farah Deeba* and S.M. Saqlan Naqvi*
HIGH-THROUGHPUT AGROBACTERIUM-MEDIATED TRANSFORMATION OF MEDICAGO TRUNCATULA IN COMPARISON TO TWO EXPRESSION VECTORS
...vector was compared with expression vector from pCAMBIA series over-expressing same gene (pCOsRGLP1). A lower number of explants generated hygromycin resistant plantlet for instance, 18.3 with pGOsRGLP1 vector as compared to 35.5% with pCOsRGLP1 vector. Transformation efficiency of PCR positive plants generated was 9.4% for pGOsRGLP1 while 21.6% for pCOsRGLP1. Furthermore 24.4% of explants -1 generated antibiotic resistant plantlet on 20 mgl of hygromycin whic...
Basit Zeshan1,2,*, Mushtaq A. Saleem2,Javed Iqbal Wattoo2, Mohd Mokhtar Arshad1 and Maizan Mohamed1
... cloned into prokaryotic expression vector resulting pET-Sf200 and confirmed the construct by sequencing. The recombinant plasmid was identified by restriction enzyme and sequencing analysis. The in vitro expression of the truncated protein was analyzed in E. coli with a molecular weight of 38kDa determined through SDS-PAGE and confirmed by Western blotting. The recombinant truncated protein was then purified from the culture med...
Aisha Khalid1, Muhammad Tayyab1,*, Abu Saeed Hashmi1, Tahir Yaqub2, Ali Raza Awan1, Muhammad Wasim1, Shagufta Saeed1, Sehrish Firyal1 and Abdul Rauf Shakoori3
...ssion host and pET28a as expression vector. Effect of various concentration of Isopropyl β-D-1-thiogalactopyranoside (IPTG), post induction time, effect of temperature and pH were examined for the maximal production of recombinant cellulase. The effect of supplementation of LB medium with additional carbon and nitrogen sources was also analyzed for maximal production of recombinant protein. Higher level enzyme activity was recorded at 25°C, pH 7.0 whe...
Gul Afshan1,2,*, Soumble Zulfiqar2, Sumaira Mehboob2, Muhammad Tahir Javed Khan1 and Abdul Rauf Shakoori2,*
...r pTG19 and subcloned in expression vector pET21a. E2 protein, expressed in insoluble form, was purified by repeated sonications, followed by denaturation and refolding through fractional dialysis with urea. Multiple alignment with other sequences showed nucleotide variations of HCV3a E2 compared with already reported sequences. A total of 140 (11%) point mutations were found in Pakistan’s gene sequence, out of which general and specific differences were...
Tingting Li, Shuangshuang Geng, Long Xie, Huiyan Xu, Aolin Luo, Pengwei Zhao, Huan Yang, XingWei Liang, YangQing Lu, XiaoGan Yang* and KeHuan Lu*
... the PiggyBac eukaryotic expression vector, which was stably expressed in Sertoli cells, thus establishing a PB-GDNF-Sertoli cell line. Quantitative real-time polymerase chain reaction (qRT-PCR) results showed that the GDNF gene expression in the PB-GDNF-Sertoli cell line was significantly increased compared with that in the control Sertoli cell line (p<0.01). Enzyme-linked immunosorbent assay (ELISA) results showed that GDNF gene secretion was also signifi...
Zengwen Huang1,2, WuReliHazi Hazihan2*, Baheti Bodai2, Kadyken Rizabek3, Nuralieva Ulzhan3, Omarova Karlygash4, Juan Zhang1 and Yaling Gu1*
...ed. The eukaryotic siRNA expression vector was constructed with the cloned gene as the target and validated at the cellular level.Three RNA interference fragments at different sites were designed based on obtained cDNA of the target gene and the siRNA expression vector pGenesil 10-3p-siRNA was constructed. The siRNA expression vector was digested, identified and sequenced, and then transfe...
