Abdel-Salam Mohamed S, Ameen Hoda H, Kassab Abdallah S M, Mahgoob Ahmed E A, Elkelany Usama S
Microbial Genetics Department, National Research Centre, 33 Albohouth St., (Formerly Altahrir St.,) Dokki, Giza, Egypt.
Department of Plant Pathology, National Research Centre, 33 Albohouth St., (Formerly Altahrir St.,) Dokki, Giza, Egypt.
J Genet Eng Biotechnol. 2018 Dec;16(2):305-310. doi: 10.1016/j.jgeb.2018.05.006. Epub 2018 May 23.
A gene encoding chitinase from has been isolated after optimization of PCR conditions. It was cloned with two different prometers, T7 promoter of the pJET1.2/blunt cloning vector and the SP6 promoter of pGEM®-T Easy vector. After transforming DH5α, two transformants were selected, CHI-NRC-4 from the first vector and T-CHI-NRC-6 from the second vector, and used for further studies. The complete CDS sequence of gene was determined and submitted to GenBank with the accession number KX268692.1. Culture supernatants of (CHI-NRC-4) and (T-CHI-NRC-6) were investigated for their inhibitory effect on egg hatch under laboratory conditions. Result showed up to 96% inhibition in egg hatching due to both transformants as compared to control which reflect the same expression efficiency of both used prometers. A greenhouse experiment was carried out to evaluate the nematicidal effect of culture supernatants of the two transformts (CHI-NRC-4) and (T-CHI-NRC-6) against infected eggplant. Obtained results showed a significant reduction in nematode population in soil and roots and enhancement in eggplant growth parameters as compared to control.
在优化PCR条件后，已从[具体来源]中分离出一种编码几丁质酶的基因。它用两种不同的启动子进行克隆，即pJET1.2/blunt克隆载体的T7启动子和pGEM®-T Easy载体的SP6启动子。转化DH5α后，挑选出两个转化体，第一个载体的CHI-NRC-4和第二个载体的T-CHI-NRC-6，并用于进一步研究。测定了[具体基因]的完整CDS序列，并提交至GenBank，登录号为KX268692.1。在实验室条件下，研究了具体菌株和具体菌株的培养上清液对[某种线虫]卵孵化的抑制作用。结果表明，与对照相比，两种转化体对卵孵化的抑制率高达96%，这反映了所使用的两种启动子具有相同的表达效率。进行了一项温室试验，以评估两种转化体(CHI-NRC-4)和(T-CHI-NRC-6)的培养上清液对感染[某种线虫]的茄子的杀线虫效果。与对照相比，获得的结果表明土壤和根中线虫数量显著减少，茄子生长参数得到提高。
