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THE ANTISENSE C-HA-RAS OLIGODEOXY-NUCLEOTIDES INHIBIT THE PROLIFERATION OF THE GASTROCARCINOMA-DNA-TRANSFORMED RAT 3-3 CELLS AND REDUCE THE P21 EXPRESSION OF THE CELLS

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Science China Chemistry 1991年第1期34卷 35-41页

作者：许毅邓国仁梁亚云李赤 Biochemistry Department Beijing Institute for Cancer Research Beijing 100034 PRC

The Rat 3-3 is a secondary transformant of the rat fibroblast cell line (Rat-1) transfectedwith total DNA of a gastrocarcinoma cell line BGC-823. The cells over-express the c-Ha-rasoncogene which contains point mutation at the 12th codon. In order to determine how theactivated c-Ha-ras oncogene expression governs the cell's transformation, two pendadecadeoxy-nucleotides AS-1 and AS-2 were synthesized. AS-1 was complementary to the single strandof the first three codons and the upstream sequence close to the ribosome binding site of c-Ha-ras mRNA. AS-2 was complementary to the 3' end of the first intron and the 5' end ofthe second exon of c-Ha-ras unripe RNA enclosed in the nucleus. The oligonucleotides couldblock either the translation of c-Ha-ras mRNA or the splicing of c-Ha-ras unripe RNA, thusinhibiting the expression of the activated c-Ha-ras oncogene and the proliferation of thetransformed cells Rat 3-3. The inhibitory effect increased with a growing concentration ofthe antisense oligodeoxynucleotide (from 4- 10 μmol/L) and reached its peak at 12 h afterRat 3-3 was treated with AS-1 and AS-2. This effect became weaker afterwards. The p21level, product of c-Ha-ras, in the Rat 3-3, was examined by ELISA method. The resultshows that the amount of p21 in the growth-inhibited cellsis about 30% of that of the controlcells.

关键词： antisense oligodeoxynucleotides transformants inhibition of cell proliferation oncogene

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Development of an efficient regeneration and agrobacterium tumefaciens mediated transformation system in Fusarium proliferatum

Development of an efficient regeneration and agrobacterium t...

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中国植物病理学会2015年学术年会

作者：刘力伟 LIU Xin LIU Li-yuan GUO Yong-bin 王树桐 College of Plant Protection Hebei Agricultural University

In apple,replant disease is widespread and has been documented in all of the major fruit-growing regions of the *** was causing serious economic losses to farmers and it also became a great hinder against the sustainable development of apple *** our previous studies,Fusarium proliferatum was approved recently to be one of the major causal agents of apple replant disease（ARD）in *** the pathogenic mechanism of *** need to be further *** this article,we report Agrobacterium tumefaciens mediated transformation（ATMT） of *** transformation efficiency of approximately 100 transformants/10 conidia,and a library containing more than 2 000 transformants was generated with the *** strain AGL-1 carrying *** tested transformants were mitotically *** hundred percent hph（hygromycin B phosphotransferase） integration into *** was identified by *** with wild-type strains,most of the mutants did not changed their colony morphologies and growth rates,only minorities of the mutants varied in their characteristics,in which25%strains accelerated their growth rate distinctly,2%strains slowed down their growth rate obviously,about 40%strains changed the number of conidia,3 strains weakened its pathogenicity,accounting 3.0%in all selected *** this study,we established genetic transformation system and T-DNA insertion mutant library of Fusarium proliferatum *** our knowledge,this is the first report of ATMT of ***,this library has been used to identify genes involved in the virulence of the pathogen,and the transformation system may also be useful to the transformation of other species of the genus Fusarium.

关键词： Fusarium proliferatum ATMT transformants

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