Roopa G, Madhusudhan M C, Sunil K C R, Lisa N, Calvin R, Poornima R, Zeinab N, Kini K R, Prakash H S, Geetha N
Eco-Biotech Laboratory, DOS in Biotechnology, University of Mysore, Manasagangotri, Mysuru 570006, Karnataka, India.
DOS in Biotechnology, Manasagangotri, University of Mysore, Mysuru 570006, India.
J Genet Eng Biotechnol. 2015 Dec;13(2):119-127. doi: 10.1016/j.jgeb.2015.09.002. Epub 2015 Oct 1.
The most promising anti-tumor agent developed in the past three decades is Taxol. It is proven to be effective against many cancers. It is necessary to isolate pharmacologically potent endophytic microbial strains from medicinal plants with special reference to Taxol production. In the current study, endophytic fungi were isolated from the bark of the medicinal plant, The isolated endophytes were identified morphologically, and further characterized by ITS-PCR using genomic DNA samples, later the products were sequenced for identification and phylogenetic linkage mapping. The samples were screened for the potential to produce Taxol or taxanes, employing PCR. The resulted data have been sequenced to confirm the presence of the two genes implicated in Taxol biosynthesis, 10-deacetylbaccatin III-10-O-acetyl transferase (DBAT) and C-13 phenylpropanoid side chain-CoA acyltransferase (BAPT). Seven samples showed the amplicons of DBAT gene and one showed the amplicons of BAPT gene. Sequencing of these products was carried out, of which one sample has revealed sequence homology to the original DBAT gene from . The present work confirms and substantiates the potential of genomic mining approach to discover novel Taxol-producing endophytic fungi.
过去三十年中研发出的最具前景的抗肿瘤药物是紫杉醇。事实证明它对多种癌症都有效。有必要从药用植物中分离出具有药理活性的内生微生物菌株，尤其关注紫杉醇的产生。在当前研究中，从药用植物的树皮中分离出内生真菌，对分离出的内生菌进行形态学鉴定，并使用基因组DNA样本通过ITS-PCR进一步表征，随后对产物进行测序以进行鉴定和系统发育连锁图谱绘制。采用PCR筛选样本中产生紫杉醇或紫杉烷的潜力。对所得数据进行测序，以确认参与紫杉醇生物合成的两个基因，10-去乙酰巴卡亭III-10-O-乙酰转移酶（DBAT）和C-13苯丙烷侧链-CoA酰基转移酶（BAPT）的存在。七个样本显示出DBAT基因的扩增子，一个样本显示出BAPT基因的扩增子。对这些产物进行测序，其中一个样本显示出与来自……的原始DBAT基因的序列同源性。目前的工作证实并证实了基因组挖掘方法在发现新型产生紫杉醇的内生真菌方面的潜力。
