Cloning and sequence analysis of pep-carboxylase from cassava

Cassava (Manihot esculenta Crantz) is very drought tolerant and hows high levels of photosynthesis at high temperatures. This makes cassava an important source of carbohydrates for several arid regions, especially in Africa. Some varieties perform better than others. Physiological performance, which...

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Autores principales: Tenjo F, FA, Mayer, Jorge Edgard
Formato: Conference Paper
Lenguaje:Inglés
Publicado: International Center for Tropical Agriculture 1993
Materias:
Acceso en línea:https://hdl.handle.net/10568/55693
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author Tenjo F, FA
Mayer, Jorge Edgard
author_browse Mayer, Jorge Edgard
Tenjo F, FA
author_facet Tenjo F, FA
Mayer, Jorge Edgard
author_sort Tenjo F, FA
collection Repository of Agricultural Research Outputs (CGSpace)
description Cassava (Manihot esculenta Crantz) is very drought tolerant and hows high levels of photosynthesis at high temperatures. This makes cassava an important source of carbohydrates for several arid regions, especially in Africa. Some varieties perform better than others. Physiological performance, which covers a wide range between C3 and C4 plants, could be increased through directed breeding. In order to do so, a better understanding of the underlying molecular mechanisms is needed, as to develop well defined breeding criteria and tests that could aid to that purpose. Several enzymes are involved in the fixation of CO2 in plants, phosphoenolpyruvate carboxylase playing a central role. We want to analyze the compartmentation of this and other photosynthetic enzymes between palisade and mesophyll cells by using in situ hybridization of the radiolabelled genes on histological sections. For this purpose the homologou gene must be isolated. We constructed a cassava genomic DNA library from the variety 996 CIAT in an EMBL 3 derived bacteriophage cloning vector. The library was screened with a maize ppc probe (obtained from T. Nelson, Yale), and after three rounds of purification, a putative cassava ppc clone of 10 kb was obtained. We are in the process of mapping and sequencing our clone. The next steps will include carrying out Northern analysis to assess levels of expression in different varieties, carrying out relatedness studies with other sequence characterized ppc genes, and the establishment of in situ hybridization techniques.
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spelling CGSpace556932024-01-09T09:49:23Z Cloning and sequence analysis of pep-carboxylase from cassava Tenjo F, FA Mayer, Jorge Edgard manihot esculenta phosphoenolpyruvate carboxylase molecular cloning analytical methods fosfenolpiruvato carboxilasa clonación molecular técnicas analíticas Cassava (Manihot esculenta Crantz) is very drought tolerant and hows high levels of photosynthesis at high temperatures. This makes cassava an important source of carbohydrates for several arid regions, especially in Africa. Some varieties perform better than others. Physiological performance, which covers a wide range between C3 and C4 plants, could be increased through directed breeding. In order to do so, a better understanding of the underlying molecular mechanisms is needed, as to develop well defined breeding criteria and tests that could aid to that purpose. Several enzymes are involved in the fixation of CO2 in plants, phosphoenolpyruvate carboxylase playing a central role. We want to analyze the compartmentation of this and other photosynthetic enzymes between palisade and mesophyll cells by using in situ hybridization of the radiolabelled genes on histological sections. For this purpose the homologou gene must be isolated. We constructed a cassava genomic DNA library from the variety 996 CIAT in an EMBL 3 derived bacteriophage cloning vector. The library was screened with a maize ppc probe (obtained from T. Nelson, Yale), and after three rounds of purification, a putative cassava ppc clone of 10 kb was obtained. We are in the process of mapping and sequencing our clone. The next steps will include carrying out Northern analysis to assess levels of expression in different varieties, carrying out relatedness studies with other sequence characterized ppc genes, and the establishment of in situ hybridization techniques. 1993 2015-01-28T14:21:42Z 2015-01-28T14:21:42Z Conference Paper https://hdl.handle.net/10568/55693 en Open Access International Center for Tropical Agriculture Tenjo F., Fernando Alberto; Mayer, Jorge Edgard. 1993. Cloning and sequence analysis of pep-carboxylase from cassava. In: Roca, William M.; Thro, Ann Marie (eds.). International Scientific Meeting Cassava Biotechnology Network (1, 1992, Cartagena de Indias, Colombia). Proceedings. Centro Internacional de Agricultura Tropical (CIAT), Cali, CO. p. 331-334. (Working document no. 123)
spellingShingle manihot esculenta
phosphoenolpyruvate carboxylase
molecular cloning
analytical methods
fosfenolpiruvato carboxilasa
clonación molecular
técnicas analíticas
Tenjo F, FA
Mayer, Jorge Edgard
Cloning and sequence analysis of pep-carboxylase from cassava
title Cloning and sequence analysis of pep-carboxylase from cassava
title_full Cloning and sequence analysis of pep-carboxylase from cassava
title_fullStr Cloning and sequence analysis of pep-carboxylase from cassava
title_full_unstemmed Cloning and sequence analysis of pep-carboxylase from cassava
title_short Cloning and sequence analysis of pep-carboxylase from cassava
title_sort cloning and sequence analysis of pep carboxylase from cassava
topic manihot esculenta
phosphoenolpyruvate carboxylase
molecular cloning
analytical methods
fosfenolpiruvato carboxilasa
clonación molecular
técnicas analíticas
url https://hdl.handle.net/10568/55693
work_keys_str_mv AT tenjoffa cloningandsequenceanalysisofpepcarboxylasefromcassava
AT mayerjorgeedgard cloningandsequenceanalysisofpepcarboxylasefromcassava