The back - exchange process is actually complex and deserving of analysis in a sequence - specific manner 返交換實際上很復雜,需要進行序列特異性的分析。
The mechanism is that the introduced complementary oligonucleotides can bind to the corresponding mrna or double - stranded dna in genome and form partial double - stranded molecules or triple - stranded nucleic acid molecules by sequence - specific and nonsequence - specific antisense action , thus the target gene will be orientationally blocked and expression of the target inhibited so that therapeutic effect could be attained . in this study , we designed a fragment of human c ii ta cdna in antisense orientation using mrna of c ii ta as template . the primers were designed based on 94 - 500 nucleotides segment in 5 " end of ciita gene so that the interested gene contained 407 base pairs which included two aug codons in 1 16 and 188 nucleotides as well as the splicing site between the first and the second exons 本研究設計以c tamrna為模板的反義cdna片段,從c ta基因5 ’端第94位到500位核苷酸段設計引物,目的片段407bp ,覆蓋第116和188位兩個aug密碼子,也包含了第一外顯子和第二外顯子間的剪接位點:用常規分子生物學方法構建了反義片段的腺病毒表達載體( padeasy - 1系統) ;腺病毒載體經hek293細胞包裝產生含反義片段的重組腺病毒,用氯化銫密度梯度離心法獲得純化的高滴度腺病毒;進行體外基因轉移,分別用反義片段真核表達載體轉染p388d1細胞和用重組腺病毒感染hela細胞,觀察導入的c ta基因反義rna抑制細胞內組成型或誘導型c ta基因表達的作用,從而達到調控mhc -類分子表達的目的。
In this paper , a series of transition metal complexes were synthesized ; interaction and reaction mechanism between complexes and dna were studied . in addition , we synthesized and analyzed a sequence - specific cleavage reagent , in which oligodeoxynucleotide was recognizing group . detailed work mentioned below were carried out : 1 本文在查閱大量文獻的基礎上,以1 , 10 -鄰菲咯啉和1 , 10 -鄰菲咯啉- 5 , 6二酮為配體合成了一系列過渡金屬配合物,詳細研究了它們與pbr322dna的相互作用及其機理。