[Expression and immunization testing of fusion protein of Newcastle disease virus in leaf tissue of transgenic rice].

Zhen-Quan Yang, Qiao-Quan Liu, Heng-Xiu Yu, Zhi-Ming Pan, Xin-An Jiao
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Abstract

Transgenic plant is an attractive bioreactor to produce recombinant protein, such as safe and economic vaccine. In present study, a plant expression binary vector pUNDV, containing the 1.7 kb fusion protein gene of Newcastle Disease Virus (NDV F) under the control of maize ubiquitin (Ubi) promoter and nos terminator was constructed, in which, the Hygromycin phosphotransferase (HPT) gene was used as the selectable marker. The expression cassette was introduced into a japonica rice variety by Agrobacterium-mediated transformation, and 6 independent transgenic rice lines were regenerated and verified by hygromycin resistance selection. The integration of target gene in transgenic plants was confirmed by PCR analysis. The results from ELISA and Western blot analyses revealed that NDV F could be expressed in the leaf of several transgenic plants. The total salt soluble proteins were extracted from leaf of transgenic plant F5, which contained the highest expressing level of NDV F protein, and used in immunization of BALB/c mice. The specific antibodies could be elicited in mice immunized with NDV F protein expressed in transgenic rice.

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[转基因水稻叶片组织中新城疫病毒融合蛋白的表达及免疫试验]。
转基因植物是生产安全、经济的重组蛋白疫苗等有吸引力的生物反应器。本研究以湿霉素磷酸转移酶(HPT)基因为选择标记,构建了含有新城疫病毒(NDV F) 1.7 kb融合蛋白基因的植物双表达载体pUNDV,该载体在玉米泛素(Ubi)启动子和nos终止子的控制下表达。通过农杆菌介导的转化,将该表达盒导入一个粳稻品种,再生出6个独立的转基因水稻品系,并进行抗潮霉素选择验证。目的基因在转基因植物中的整合通过PCR分析得到证实。ELISA和Western blot分析结果表明,NDV F可以在几种转基因植物的叶片中表达。从NDV F蛋白表达量最高的转基因植物F5叶片中提取盐溶性总蛋白,用于BALB/c小鼠免疫。用转基因水稻表达的NDV F蛋白免疫小鼠,可诱导出特异性抗体。
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[Influence of B mating-type factor on recovery of nuclear types from dikaryons in Lentinula edodes]. [Expression and immunization testing of fusion protein of Newcastle disease virus in leaf tissue of transgenic rice]. Nuclear matrices and matrix attachment regions from Green alga: Dunaliella salina. [Molecular basic of interaction between disease resistance gene and avirulence gene]. [Progress in the study on diacylgycerol acyltransferase (DGAT)-related genes].
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