The mechanisms of gene silencing include copy numbers and configuration of transgene , integration sites in plant and transcription of transgene , etc 基因沉默的機(jī)制是多方面的,包括轉(zhuǎn)基因的拷貝數(shù)和構(gòu)型、在植物上的整合位點、轉(zhuǎn)基因的轉(zhuǎn)錄水平等。
To abolish transcription efficiency affected by copy number or integration site in chromosome , all the vectors were integrated into frt site by co - transfecting with an flp recombinase producing plasmid 應(yīng)用w七stemblot對其中的20個克隆進(jìn)行檢測,選出一個caspase一3表達(dá)量最低的細(xì)胞克隆a6 。
In abroad , the study of integration site used for transgenic detection had just begun . in this study , according to the collection of the global commercialized transgenic crops , select seven exogenous genes which basically cover the total commercialized crops , namely camv35s and fmv promoter , nos terminater , mark gene nptii , and aim genes pat , epsps and cryia ( b ) . use endogenous 18srrna gene as collate , design a large pairs of specific primers , screen the optimum primers groups , optimized the test condition and parameters , establishing the qualitative pcr detection system 本研究根據(jù)收集的國內(nèi)外已商品化的轉(zhuǎn)基因作物品種,選擇了能基本覆蓋商品化轉(zhuǎn)基因品種的7個外源基因,即: camv35s 、 fmv啟動子、 nos終止子、 npt標(biāo)記基因和目的基因pat 、 epsps 、 cryia ( b )作為篩選目標(biāo),以植物18srrna基因作為內(nèi)源參照基因,設(shè)計了多對特異性引物,并篩選出最佳組合,優(yōu)化了檢測條件和參數(shù),建立了pcr定性檢測方法體系。
Transgenic integration site detection is the efficient method to identify whether the transgenic crops are approved to import or export . some quantitative detection methods were reported in 2000 and 2001 , and that is about bt11 and mon810 corn . real - time 5 " - nuclease pcr had been previously used successfully to determine quantitatively roundup ready ( rr ) soy and btl76 maize in food 轉(zhuǎn)基因整合位點檢測是鑒定轉(zhuǎn)基因品種是否為批準(zhǔn)進(jìn)口轉(zhuǎn)基因作物的有效的特異性方法,國外在2000年和2001年已開展了轉(zhuǎn)基因玉米bt11 、 mon810兩個品種整合位點的特異性檢測方法研究。
The env protein deduced from env gene encodes the hydrophilic surface protein ( su ) and the hydrophobic transmembrane domain ( tm ) that determine the specific interaction between virus particles and cell surface receptors during retroviral entry . the su of retroviruses is a highly variable genetic element , containing receptor binding sites and major antigenic determinants . exjsrv - specific dna probes were derived . by using these dna probes in tissue hybridization . we successfully identified jsrv mrna expression and proviruses dna in sheep lung tissues infected with jsrv and control group has no postive signals , validating the use of exogenous virus - specific dna probes in the analysis of oncogenic proviral integration sites and identification of integrated exogenous proviral sequences 用地高辛隨機(jī)引物法標(biāo)記exjsrv特異的env片段,制備探針,原位雜交檢測spa肺組織中的rna及前病毒dna ,結(jié)果表明spa患羊肺組織內(nèi)有jsrvenv基因mrna的表達(dá),同時也檢測到了前病毒dna ,而相應(yīng)的陰性對照卻無陽性信號,證實外源性病毒特異的dna探針在致瘤性前病毒的整合位點和整合的外源性前病毒的檢測中具有可信度。