NICOTIANA TABACUM AS A POTENTIAL PLATFORM FOR THE PRODUCTION OF RECOMBINANT ANTI-TOXOPLASMA SINGLE-CHAIN VARIABLE FRAGMENT (scFv) ANTIBODY

• DOI: 10.22452/mjs.vol39no2.1
• Publication Date: 2020-07-07T07:49:47Z
• Authors: Fatin Iffah Rasyi...; Pei See Go; Boon Chin Tan; Nur Ardiyana Rejab; Rofina Yasmin Othman; Norzulaani Khalid
• Keywords: Single-chain variable fragment

Plant systems have now gained much attention as they provide low risk of pathogen contamination and cost production, appropriate post-translational modification, multimeric assembly capability compared to the mammalian bacterial hosts. In this study, we aimed produce single-chain fragment antibody (scFv) encoding anti-Toxoplasma proteins, TP60 in Nicotiana tabacum cv. SR1. Leaf explants N. were infected with Agrobacterium tumefaciens strain LBA4404 harbouring binary vector pCAMBIA1304 containing gene. Putative transformants confirmed through GUS GFP qualitative assays. Bands observed at predicted size 914 bp presence transgene was stably integrated both T0 T1 tobacco genome. FV12-6 transgenic line produced highest mRNA expression (7-fold) correlated accumulation recombinant scFv (0.52 % total soluble protein), followed by FV16-10 (0.25 %), FV17-7 FV3-11 (0.18 %). The did not affect growth plants segregation analysis FV3, FV12, FV16 generation integration within a single locus according 3:1 Mendelian’s law. These findings indicated possibility using bio-factory for protein production.   Abstrak Sistem tumbuhan kini telah mendapat banyak perhatian kerana mereka menyediakan risiko yang rendah terhadap pencemaran patogen dan kos pengeluaran, pengubahsuaian pasca-protein translasi sesuai, keupayaan penggabungan multimerik berbanding dengan tentera mamalia bakteria. Dalam kajian ini, kami berhasrat untuk menghasilkan antibodi serasi rantaian tunggal mengekodkan anti-Toxoplasma, di Eksplan daun dijangkiti oleh menyimpan vektor dedua mengandungi gen TP60. Transforman putative disahkan melalui ujian kualitatif GFP. Band-band diperhatikan pada saiz ramalan mengesahkan kehadiran transgen tersebut diintegrasi secara stabil dalam genom tembakau T1. Individu transgenik ekspresi tertinggi (7 kali ganda) berkait pengumpulan rekombinan (0.52% daripada jumlah larut), diikuti (0.25%), (0.18%). Ungkapan tidak menjejaskan pertumbuhan analisis pemisahan generasi integrasi satu lokus mengikut 3: 1 undang-undang Mendelian. Penemuan ini menunjukkan kemungkinan menggunakan sebagai kilang bio pengeluaran rekombinan. Keywords Agrobacterium-mediated transformation, plant molecular farming, protein, (scFv), tobacco, toxoplasmosis