MOLECULAR CLONING, PROKARYOTIC EXPRESSION AND PREPARATION OF ANTISERUM AGAINST LILY SYMPTOMLESS VIRUS TGB1 PROTEIN
Journal Title: JOURNAL OF ADVANCES IN AGRICULTURE - Year 2017, Vol 7, Issue 1
Abstract
The triple gene block gene TGB1 was amplified by RT-PCR from lily leaves infected with Lily symptomless virus and cloned into prokaryotic expression vector pET-28a(+). The recombinant vector was transformed into Escherichia coli strain BL21 (DE3). On induction with isopropyl β-D-1-thiogalactopyranoside,TGB1 protein was highly expressed and the molecular weight was 29 kDa (including a His-tag-containing fusion). After protein purification by Ni2+-NTA affinity chromatography, a polyclonal antibody against TGB1 was raised in mouse. Western blot analysis showed that the antiserum reacted specially with the TGB1 protein of LSV. ELISA and RT-PCR confirmed that the antiserum reacted specially with lily leaves infected with LSV, and can be used for a rapid test for LSV. The antibody produced in this work may be used for future immunohistochemistry and functional study of TGB1.
Authors and Affiliations
Pinsan Xu, Xinlei Wang, Zhengyao Zhang
Keywords
Related Articles
- EP ID EP652415
- DOI 10.24297/jaa.v7i1.5794
- Views 176
- Downloads 0
Multivariate statistical analysis of cold season wheat yield under drought conditions Alireza Razavi
Effect of drought stress on cold season wheat varieties investigated at Jolgeh-rokh station of Torbat Heydarieh agricultural and resources research center during 2012 and 2013. The experiment contained 18 inbred lines an...
Perception of Ethiopian Mango Farmers on the Pest Status and Current Management Practices for the Control of the White Mango Scale, Aulacaspis Tubercularis (Homoptera: Diaspididae)
White Mango Scale (WMS), Aulacaspis tubercularis Newstead (Hemiptera: Diaspididae), was occurred in Ethiopia in 2010. It was introduced from Asia (India) through infected mango seedlings. Currently the insect problems in...
GENETIC DIVERSITY OF CASSAVA (Manihot esculenta Crantz), (EUPHORBIACEAE) GROWN IN THREE AGROCLIMATIC ZONES OF CENTRAL AFRICAN REPUBLIC
Cassava (Manihot esculenta Krants) introduced in the Central African Republic (CAR) in the 1850s, is now the staple food of the population. It does not know the genetic diversity of cultivated accessions in peasant commu...
ACCUMULATION AND PARTITION OF Fe, Zn, Cu, Mn AND Na IN MACRO AND MICRONUTRIENT-DEFICIENT COWPEA PLANTS
Cowpea (Vigna unguiculata) is an important crop for people living in the semi-arid tropics where it is used as food, animal feed and forage. Researches related to nutritional status of plants are important, principally i...
The Stimulating Effects of Imazalil and Carbendazim Fungicides on in Vitro Propagation and ex Situ conservation of the Medicinal Balkan Range-Restricted Sideritis Raeseri Boiss & Heldr. Subsp. Raeseri
Sideritis raeseri Boiss & Heldr. subsp. raeseri, known in Greece as Mountain tea of Parnassus or Velouchi is a range restricted medicinal plant of the Balkan peninsula. Conventional propagation methods did not allow...