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Evaluation of Synthetic Epsps Gene in Genetically Transformed Plants

Thesis Info

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Author

Imran, Muhammad

Program

PhD

Institute

Pakistan Institute of Engineering and Applied Sciences

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Biotechnology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/12812/1/M.%20Imran%20Thesis%20%28PIEAS%2c%2010-7-1-059-2010.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726142648

Similar


Codon optimized synthetic epsps gene was used to develop transgenic tobacco and sugarcane lines using Agrobacterium and biolistic approach, respectively. Putative transgenic tobacco lines analyzed by qRT-PCR showed variable mRNA transcript levels. Southern hybridization performed for determining gene integration patterns showed multiple copy inserts (1-4) of transgene in various tested lines. Immunoblot strips confirmed the translation of cp4-epsps mRNA transcripts in T1 line. Malachite green assay showed variable EPSPS activity levels in various lines in response to glyphosate application. Transgenic tobacco lines exposed to 1% (v/v) glyphosate at 4 to 5 leaf stage showed variable resistance levels ranging from moderate level tolerance to high tolerance. The cp4-epsps mRNA transcript levels varied from genotype to genotype in tested transgenic sugarcane lines regardless of the integration of multiple copy number (3-6) of transgene. Genetic transformation with pyramided constructs results in the integration of multiple genes at a single locus, which is desirable in terms of expression efficiency of transgenes. The cp4-epsps gene was pyramided with insect resistant cry1Ac and cry2Ab genes and cloned in pSb187 vector was used to genetically transform tobacco using Agrobacterium mediated approach. Transgenic lines were confirmed through PCR using gene specific primers of respective genes. The mRNA transcript level of cp4-epsps gene detected through qRT-PCR using GAPDH gene as an internal control was found to be variable among different transgenic lines. Transgenic lines were tested for checking the translational efficiency of all three genes using immunoblot strips. Transgenic tobacco lines found expressing all three proteins were subjected to detached leaf insect bioassays to evaluate the effectiveness of Bt genes in putative transgenic tobacco against insect attack using first instar larvae from Spodoptera littura and Helicoverpa armigera. Transgenic tobacco found to be resistant against these insects showed that cry1Ac and cry2Ab genes were efficient enough in pyramided form. When these transgenic tobacco lines were exposed to glyphosate at 10 to 12 leaf stage, chlorotic symptoms were observed which hinted that herbicide tolerance potential of transgene declined with increasing age of the plants. In order to overcome the spatio temporal gene expression issues, a dual copy epsps expression cassette synthesized under different regulatory elements, cloned in pGreen0029 binary vector was used to generate transgenic tobacco. These tobacco lines exhibited excellent resistance against 19% (v/v) glyphosate at 10-12 leaf stage. This construct has shown tremendous potential regarding transgene expression levels which is required in the field crops. This dual cassette cp4-epsps gene has been recommended for generation of glyphosate resistant cotton and other commercially important cultivars.
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فصل چہارم: حدودوقصاص کے نفاذ میں معاون سماجی اداروں کا کردار

معاشرے کی بنیادی اکائی فردہے اور معاشرہ مختلف افراد کے میل جول ، رہن سہن ، مشترکہ مفادات اور روحانی و مادی ضروریات کے تحفظ کے لئے وجود میں آتا ہے ۔ جب معاشرے کا ہر فرد اپنی تمام ضروریا ت کی انجام دہی خود نہیں کر سکتا ،تو ریاست تمام ذمہ داریوں کو مشترکہ طور پر لوگوں کی صلاحیت ، قابلیت اور تعلیم کے مطابق تقسیم کر دیتا ہے ، جس سے معاشرے کی تمام ضروریات پوری ہوتی ہیں ۔یہ سب لوگ اجتماعی طور پر معاشرے کے تمام امور انجام دے رہے ہوتے ہیں ۔ اب اکیلا فرد تو اتنے سارےکا م سرانجام نہیں دے سکتا۔ اسی طرح معاشرے کے مقتدر ، بااختیار اور تعلیم یا فتہ اہل افراد اکٹھے مل کر اپنی خدادادصلاحیتوں کو استعمال کرتے ہوئے پورے معاشرے کی اصلاح و تعمیر کا بھی فریضہ ادا کرتے نظر آتے ہیں ، جیسا کہ اسلامی معاشرےکا قیام، اسلامی قوانین کا نفاذ ، امن وامان کا قیام ، ذرائع ابلاغ کا درست استعمال اور علمائے کرام و خانقاہی نظام کا کردار وغیرہ۔ قوانین حدودو قصاص کے نفاذ کے حوالے سے معاشرہ ریاست کے بغیر یہ قوانین خود نافذ نہیں کر سکتا ، البتہ معاشرہ اسلامی حکومت کے قیام اور معاشرتی اصلاح کے حوالے سے جدو جہد کر سکتا ہے ۔ لہذا امت مسلمہ کی اجتماعی ذمہ داریوں کی بجاآوری امت کے تمام افراد کے لئے فرض کفایہ ہے۔ درحقیقت حد ودکا نفاذ اسلامی حکومت کے قیام سے مشروط ہے اورحاکم وقت یا اس کا نمائندہ ہی حد کا نفاذ کر سکتا ہے ،جیساکہ سورۃ النور کی آیت نمبر دو کی تفسیر کرتے ہوئے علامہ قرطبی لکھتے ہیں
"الْخِطَابُ لِلْمُسْلِمِينَ، لِأَنَّ إِقَامَةَ مَرَاسِمِ الدِّينِ وَاجِبَةٌ عَلَى الْمُسْلِمِينَ، ثُمَّ الْإِمَامُ يَنُوبُ عَنْهُمْ، إِذْ لَا يُمْكِنُهُمُ الِاجْتِمَاعُ عَلَى إِقَامَةِ الْحُدُودِ." 389
"یہ خطاب تمام مسلمانوں سے ہے اس لیے...

Economic Policies of Pakistan During Military Rules an Analytical Study in Islamic Perspective

In the history of Pakistan, three military governments ruled for a quite long period. Nonetheless, they made no stable policies for the economic development of Pakistan. As a result, Pakistan could not make progress on strong foundation. Economic policies were meant for a shorter interval. Therefore, they only resulted in cosmetic change in the economy. The results of these policies were restricted to a certain class and the public suffered from the negative impact of these policies. Public welfare got secondary importance which resulted in economic inequality. Wealth circulated within a few families. In all the three reigns, no strategy was formed for the paying of external debt. On the one hand, the salvation of Pakistan was put at stake by increasing external debt and aid while on other hand, masses suffered added direct and indirect taxes. In these conditions, common man could not fulfill his basic necessities of life. Pakistan could make considerable progress, if any of the governments might have considered Islamic principles of Islam.

Exchange Rate Volatility in Developing Countries: Implications for Trade and Capital Flows in Pakistan

An analysis to investigate the impact of exchange rate volatility was performed in this study considering trade and capital flows within Pakistan`s trade partners perspectives. Two different sets of volatility variables were derived and compared by using two currencies as a base for the measurement of exchange rates, viz. US dollar and Pak-rupee, for this purpose. The Generalized Autoregressive Conditional Heteroskedasticity (GARCH) model based approach was considered appropriate to calculate and generate the variance series as a proxy of volatility. Some other important specifications like Component (CGARCH), Exponential (EGARCH), Power or Nonlinear (PGARCH), Threshold (TGARCH) models were included in analysis to capture the impact of volatility clustering, asymmetrical news, leverage effect, dominant shock, impact of bad news, elements of long run persistence and the nonlinearities in volatility of exchange rates respectively. Monthly series, beginning from January 1970 to December 2009, reflecting relatively high frequency data were used to make detailed analysis of long run and short run volatility scenarios. A Sample of only twenty-nine countries was considered after sorting out the forty selected significant trade partners of Pakistan on the basis of data availability regarding all relevant variables, with limitation that many developing countries could not be included in final analysis. The resultant sample contained fifteen developed countries that shared about fifty percent of total trade with Pakistan and only fourteen developing countries. The main objective of this study was to explore the linkages of exchange rate volatility under trade and capital flows context by comparing both identical currencies based exchange rate volatilities within short and long run scenarios primarily for Pakistan along with her counterpart developing countries. The application of panel data techniques based on fixed effects models allowed us to accomplish this objective because it ensures the removal of any undesirable distorting effects that may exist due to the omitted variable bias in the system of equations and controls the inter-country variations by cross effects and intra-country variation by period effects. Mainly GARCH (1,1) models, for both currencies exchange rates, were used to derive effective volatility proxy. Whereas, other volatility proxies as obtained through different GARCH based specifications permitted us to diagnose in detail the nature of volatility relevance implied within the context of trade and capital flows in developing countries like Pakistan. Fixed effect models, as compared to random effect models, were found more appropriate to estimate trade related functions including real trade balance, real exports, real imports, and real capital flows as well. Mainly, pooled data least squared dummy variables (LSDV) technique allowed us to develop different long run and short run fixed effect models by employing all sampled countries to analyze mutual trade aspects. The sample was further divided into two sub-groups: developed countries sub- sample and developing countries sub-sample, with the purpose to compare the impact of volatility under various trade options to evaluate developing countries trade suitability within their own group as potential strategy to avoid exchange rate volatility distortions. The results revealed that exchange rate volatility remained highly significant in short run irrespective of the currency used and lead to reduce exports as well as imports when trade was done mutually among sampled countries including both developed and developing ones. However, United States Dollar based exchange rate volatility (USD-ERV) helped to improve the trade balance while it was deteriorated with increased Pakistan Rupee based exchange rate volatility (PKR-ERV), these results remained consistent in mutual and intra developing countries both cases. Increased capital outflows were found significant independent of the currency in use across developed and developing countries when they exchanged the financial assets mutually. Some interesting findings were obtained within developing countries framework; basic volatility effect was absent for real exports with both currencies, it was absent for real imports with PKR but inverse with USD; No such effect was even existed for capital outflows in both currencies. However caused reduction in long run US dollar based imports and Pak-rupee based exports and imports both. Pak-rupee remained useful to avoid volatility impact when intra-developing countries trade occurs. However, in short run, mutual trade among developed and developing countries expanded with rise in volatility. The favorable effect of using US dollars appeared in case of trade balance only. Capital outflows increased when volatility was arrived during mutual trade irrespective of the currency in use. Effects of other volatility components on trade remained significant mostly in terms of US dollars as compared to Pak-rupee.