TITLE

Cloning, functional expression and characterization of a phytocystatin gene from jelly fig (Ficus awkeotsang Makino) achenes

AUTHOR(S)
Douglas J. H. Shyu; Yi-Mei Yong; Hsi-Chi LU; Yueh-Mei Cheng; Jason T. C. Tzen; Wing-Ming Chou
PUB. DATE
October 2011
SOURCE
Botanical Studies;2011, Vol. 52 Issue 4, p407
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
A cDNA clone encoding a phytocystatin was isolated and identified from about 300 expressed sequence tag (EST) clones in maturing jelly fig (Ficus awkeotsang Makino) achenes. This clone, named FaCYS, consists of 582 bp encoding 114 amino acids with a putative signal peptide. The predicted mature protein contains no cysteine and has a molecular mass of 10.8 kDa with an isoelectric point (pI) of 9.7. Fa- CYS constructed in nonfusion and fusion vectors were overexpressed in Escherichia coli as nonfusion and histagged recombinants, respectively. Both recombinants were found in the soluble fractions of the cell extracts. The purified nonfusion and his-tagged FaCYS exhibited papain inhibitory activity with similar Ki values of 2.7 x 10-7 M and 2.4 x 10-7 M, respectively. In addition, his-tagged recombinant proteins showed inhibitory activity toward human cathepsin B, cathepsin L and ficin with Ki values of 5.6 x 10-7 M, 3.0 x 10-8 M and 2.0 x 10-7 M, but no inhibitory activity against stem bromelain. It was tolerant at a wide range of pH values and thermally stable up to 50°C for 30 min. Furthermore, his-tagged FaCYS could arrest the fungal growth of Glomerella cingulata and Sclerotium rofsii.
ACCESSION #
69720866

 

Related Articles

  • Humanizing plantibodies. Fletcher, Liz // Nature Biotechnology;Apr2001, Vol. 19 Issue 4, p319 

    Focuses on plants as cost effective means for producing recombinant human proteins. Limitations of the therapeutic value of the plant-derived product; Creation of transgenic plants to express human genes; Resemblance in the glycosylation pattern of plant-derived antibody with mammalian antibody.

  • Molecular regulators of phosphate homeostasis in plants. Wei-Yi Lin; Shu-I Lin; Tzyy-Jen Chiou // Journal of Experimental Botany;Apr2009, Vol. 60 Issue 5, p1427 

    An appropriate cellular phosphate (Pi) concentration is indispensable for essential physiological and biochemical processes. To maintain cellular Pi homeostasis, plants have developed a series of adaptive responses to facilitate external Pi acquisition and to limit Pi consumption and to adjust...

  • Rapid and reliable extraction of genomic DNA from various wild-type and transgenic plants. Tae-Jin Kang; Moon-Sik Yang // BMC Biotechnology;2004, Vol. 4, p20 

    Background: DNA extraction methods for PCR-quality DNA from calluses and plants are not time efficient, since they require that the tissues be ground in liquid nitrogen, followed by precipitation of the DNA pellet in ethanol, washing and drying the pellet, etc. The need for a rapid and simple...

  • Two decades of plant-based candidate vaccines: a review of the chimeric protein approaches. Soria-Guerra, Ruth; Moreno-Fierros, Leticia; Rosales-Mendoza, Sergio // Plant Cell Reports;Aug2011, Vol. 30 Issue 8, p1367 

    Genetic engineering revolutionized the concept of traditional vaccines since subunit vaccines became reality. Additionally, over the past two decades plant-derived antigens have been studied as potential vaccines with several advantages, including low cost and convenient administration. More...

  • A Novel 5-Enolpyruvylshikimate-3-Phosphate Synthase Shows High Glyphosate Tolerance in Escherichia coli and Tobacco Plants. Gaoyi Cao; Yunjun Liu; Shengxue Zhang; Xuewen Yang; Rongrong Chen; Yuwen Zhang; Wei Lu; Yan Liu; Jianhua Wang; Min Lin; Guoying Wang // PLoS ONE;Jun2012, Vol. 7 Issue 6, p1 

    A key enzyme in the shikimate pathway, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) is the primary target of the broad-spectrum herbicide glyphosate. Identification of new aroA genes coding for EPSPS with a high level of glyphosate tolerance is essential for the development of...

  • Delivery of multiple transgenes to plant cells by an improved version of MultiRound Gateway technology. Buntru, Matthias; Gärtner, Stefanie; Staib, Lena; Kreuzaler, Fritz; Schlaich, Nikolaus // Transgenic Research;Feb2013, Vol. 22 Issue 1, p153 

    At present, only few methods for the effective assembly of multigene constructs have been described. Here we present an improved version of the MultiRound Gateway technology, which facilitates plant multigene transformation. The system consists of two attL-flanked entry vectors, which contain an...

  • A Simple, Chisel-Assisted Mechanical Microdissection System for Harvesting Homogenous Plant Tissue Suitable for the Analysis of Nucleic Acids and Proteins. Brandt, Stephan; Walz, Christina; Schad, Martina; Pavlovic, Nada; Kehr, Julia // Plant Molecular Biology Reporter;Dec2003, Vol. 21 Issue 4, p417 

    Many physiological processes are limited to specific tissues or even specific cell types. Analysing entire plants or organs results in averaged data of all cell types contained in the sample; thus, specific metabolic functions cannot be assigned to individual cell types. A higher spatial...

  • RNA Isolation From High-Phenolic Freeze-Dried Tea (Camellia sinensis) Leaves. Jaiprakash, Maya Rani; Pillai, Beena; Venkatesh, Purna; N., Subramanian; Sinkar, Vilas P.; Sadhale, Parag P. // Plant Molecular Biology Reporter;Dec2003, Vol. 21 Issue 4, p465 

    With minor modifications, we applied a previously reported RNA isolation protocol that used guanidine hydrochloride to leaves of lyophilized (freeze-dried) tea (Camellia sinensis). Plant tissue must be preserved in its collected state, especially when genome-wide expression profiles are studied....

  • DNA Isolation From Fresh, Dry Plant Samples With Highly Acidic Tissue Extracts. Warude, Dnyaneshwar; Chavan, Preeti; Joshi, Kalpana; Patwardhan, Bhushan // Plant Molecular Biology Reporter;Dec2003, Vol. 21 Issue 4, p467 

    Current DNA isolation methods are limited in their ability to obtain quality and/or quantity DNA from plants, such as Emblica officinalis, Terminalia belerica, and Terminalia chebula, which have low pH and high amounts of secondary metabolites in tissue extracts. Our modified DNA isolation...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics