71. Nuh/Prophet Noah
I/We begin by the Blessed Name of Allah
The Immensely Merciful to all, The Infinitely Compassionate to everyone.
71:01
a. Truly, WE assigned Noah with a Divine Mission to his people, saying:
b. ‘Warn your people of the consequences of disobedience to the Divine Message before a grievous punishment would actually afflict them!’
71:02
a. Thus he proclaimed:
b. ‘O My People!
c. I have been assigned to you as a Clear Warner and my message is simple to understand and easy to follow.
71:03
a. The message is:
b. ‘Submit in worship and reverence to Allah alone, and
c. fear HIS disobedience and be reverent to HIM,
d. and, thus, follow me’ in guidance.
71:04
a. By so doing, ‘HE will be forgiving some of your past sinful offenses, and grant you respite till an appointed time - the time of death – instead of destroying you immediately.
b. But when Allah’s Appointed Time will come, then it would neither be averted nor delayed, if only you knew.’
71:05
a. So Noah spent hundreds of years conveying the Divine Message but he faced immense opposition, insult, and mockery.
b. Thus he submitted:
c. O ‘My Rabb - The Lord!
d. Truly, I have appealed to my people by night and by day, as I was assigned to do,
71:06
a. but the more I would call, the farther they would run away’ from my advocacy.
71:07
a. And, truly, each time I would call them so that they would accept The Truth and YOU may
thus grant forgiveness to them,
682 Surah 71 * Nuh
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Allah has revealed Holy Quran to guide and transform the lives of human being. According to Hadith, Quran was revealed in seven dialects because it is the name of wisdom. So, seven alphabets have numerous philosophies and benefits embedded in them. In this article introduction of Quran along with literal and figurative meanings have been elaborated. Details of seven alphabets have been explained vividly besides literal and figurative meanings of seven alphabets have been expounded. After that three important axioms of savants have been narrated. Amongst them, Imam Razi’s axiom is cited specially. In the last, modern axiom is given with explanation in a lucid way. A part from that, the logics and reasons behind the revelation of Quran in seven dialects have been deliberated including revelation of Quran in Arabic language, affection of the Holy Prophet (P.B.U.H) for Ummah, convenience for Muslim Nation, satisfaction for nature, eradication of linguistic bias amongst Arabs, consensus of two commands, narration of two commands of different versions. Abundance of virtues, legitimacy of Qur’an, statures of readers and replicators of readers and others have been mooted exhaustively. In the end, article is summarized in the light of modern era.
The aim of present study is the cloning of F and HN gene in plant expression vector to develop plant-based edible vaccine against Newcastle disease virus (NDV) of poultry. NDV was collected from Veterinary Research Institute, Lahore, Pakistan on request. ~1662bp F gene and ~1712bp HN gene were PCR amplified from cDNA of NDV that were TA cloned and further evaluated through sequencing. BLAST results determined the specificity of these genes in accordance with previously reported Mukteswar strain (Accession # GU182327). Both F and HN genes were ligated into pET30a expression vector to produce recombinant pET-F and pET-HN. Both recombinant constructs were transformed in E. coli Rosetta cells to study the prokaryotic expression of immunogenic protein. Induction by adding IPTG generated increased yield of protein and SDS polyacrylamide gel electrophoresis confirmed protein on desired size (~67kDa for F and ~69kDa for HN). Further western blot analysis, confirmed specificity of protein through antigen antibody reaction at proper size. Protein purification using IMAC affinity chromatography was performed and the appearance of single band of F protein at ~67kDa and HN protein at ~69kDa confirmed the specificity of our desired immunogenic protein. 2D and 3D structural analysis of F and HN proteins through Immune epitope database (IEDB) analysis resource tool revealed that more than 70% of its sequence is antigenically active and the predicted protein regions behave as epitopes. After prokaryotic expression of both F and HN genes, the next major objective of this study was to construct pCAMBIA 1301 with both F and HN for plant transformation. After confirming F and HN inserts, recombinant pCAMBIA-F+HN plasmid was electroporated into Agrobacterium cells (LBA4404) using a Bio-Rad electroporation device. After confirming the presence of F or HN genes in Agrobacterium, embryos from inbred lines of maize were transformed with recombinant pCAMBIA-F+HN by Agrobacterium mediated nuclear transformation. Further, the presence of F and HN genes in transgenic maize plants were confirmed through different molecular biological tools. The putative plants were confirmed through polymerase chain reaction (PCR). PCR confirmed a short fragment of ~181bp and ~191 for F and HN genes respectively. RT-PCR analysis confirmed the expression of F and HN genes in corn leaves and seeds respectively. The comparitive analysis of Ct values obtained by qRT-PCR revealed that the expression of F gene increased from 2-7.1 fold. Similarly, comparison of Ct value by qRTPCR confirmed that expression of HN gene increased from 0.5 to 4 fold as compared to negative control.Protein expression of F and HN genes were confirmed through ELISA and western blot by using gene specific antibodies. In ELISA, both F and HN proteins expression were observed in putative plants. The maximum obtained concentration of F gene was in the range of 0.15μg/ml to 0.166 μg/ml. Similarly, maximum obtained concentration of HN protein wasin the range of 0.195μg/ml 0.24μg/ml. The plants with high mRNA expression of F and HN genes were confirmed through western blot analysis. Transgenic plants produced a fragment of 67kDa for F protein and 69kDA for HN protein which confirmed the expression of transgene. Furthermore, immuniztion of chicks with transgenic maize and immune response generated by ELISA results showed production of anti-NDV antibodies in sera of chicks. On the other side ELISA results from the sera of chicks having non-transgenic diet did not induce any significant immune response. This is a key achievement of this study, which can lead towards development of plantbased edible vaccine against Newcastle disease virus of poultry.