质粒体

Wtp53 has obvious antiblastic effect towards Y79 cells, and the apoptosis rate of Y79 cells was the highest after transfection mediated by ultrasound-microbubble.

结论wtp53基因对RB瘤细胞具有较明显的抑制生长的作用；质粒+微泡+超声辐照，质粒+脂质体转染，以及质粒+超声辐照，均可促进Y79细胞的凋亡，但超声微泡介导的转染引起的凋亡率高于脂质体转染，而二者的凋亡率又明显高于质粒+超声转染。

After thermal induction, no specific recombinant protein band in SDS-PAGE was found, but G-CSF activity was detectable. Therefore, a new recombinant plasmid pBVHG2 expressing hG-CSF hybrid protein with additional 8 amino acids which could be cut off specifically with the help of mucosal enterokinase at the N terminus of hG-CSF was constructed.

含此质粒菌株虽然表达菌体裂解后可测得明显的生物学活性，但SDS-PAGE仍未见特异表达产物带；因此，再应用相同方法，在hG-CSF cDNA突变体5′端增加24核苷酸对的FLAG肽编码序列，构建了hG-CSF杂交蛋白（hG-CSF天然蛋白N末端增加8aa的FLAG肽，后者可由肠激肽酶切除）的表达质粒pBVHG2。

A pair of primers that contained flanking hydropathic amino acid codons were synthesized, to amply the sequence coding for 10~34 aa in the precursor sequence. The amplified fragment was inserted into pRSET-A to generate the first double repeat of the precursor gene. By utilizing a pair of isocaudamer BamH I and Bgl II sites, and another downstream Hind III site of plasmid pRSET-A, following a series of simple double digestions and ligation of the resulted products, a series of repeat (3, 4 and 6) precursor peptide fragment genes were derived.

本实验设计一对两侧含编码疏水性氨基酸密码子的引物，经过扩增前导序列10~34aa基因序列，并重新克隆入质粒pRSET-A构建串联二聚体后，再利用质粒pRSET-A的BamH I / Bgl II同尾酶克隆位点，经一系列简单的酶切和连接，快速构建这一前导肽中不含组氨酸标签序列的串联多聚体基因，并成功表达其六聚体重组蛋白。

Methods: Two recombinant palter plasmid containing whole CDNA sequences of HMCD59 transfected CHO cells by lipofectamine with PCDNA respectively. Then G418 was used to select positive clone.

分别构建两种含有HMCD59全长CDNA序列的重组pALTER质粒，并与pCDNA3质粒，按3：1比例混合后，运用脂质体介导法共转染CHO细胞(编号为HM5-CHO及HM6-CHO)，用新霉素类似物G418初步筛选出阳性克隆。

Recombinant plasmid pSVH 7 DNA of avian influenza virus H7 subtype heamagglutinin gene was encapsulated with DC-chol/DOPE liposomes and PC/chol/SA liposomes separately. Two-week old SPF chickens were intramuscularly inoculated with 50 μ g/0.2ml of the liposome entrapped PSVH 7 DNA. Four-weeks later, each chicken was challenged with 0.1ml 〓 AIV . One week after the challenge, the secretion of the cloacas was collected and transfected to chicken embryos to isolate the virus. The virus was isolated from 6/6 of the control group, 1/6 of the naked DNA group, 1/6 of the PC/chol/SA entrapped DNA group and 0/6 of the DC-chol/DOPE liposome entrapped group. The HI antibody titers (log2) of the four groups were 6. 83±0.98, 7. 0±1. 26, 7. 83±1. 17 and 8. 00±0.89 respectively 1-week after challenge, and 8. 5±0.55, 8. 17±0.82, 8. 68±0.45 and 9. 33±0.54 respectively 2-week after challenge. The results showed that inoculation of liposome entrapped DNA significantly enhanced resistance to virosis in animals.

将含禽流感病毒H7亚型血凝素基因的重组质粒pSVH7用DC-chol阳离子脂质体和胆固醇／卵磷脂／十八胺脂质体包裹，免疫2周龄SPF鸡，4周后用同型禽流感病毒进行人工感染，1周后采集泄殖腔分泌物分离病毒，结果未免疫组6／6分离到病毒，裸质粒DNA免疫组1／6分离到病毒胆固醇／卵磷脂／十八胺脂质体包裹DNA免疫组1／6分离到病毒，DC-chol脂质体DNA免疫组没有分离到病毒（0／6）：人工感染后1周各组的HI抗体（Log2）分别为6.38±0.98，7.00±1.26，7.83±1.17，8.00±0.89，2周后为8.50±0.55，8.67±0.82，8.68±0.45，9.33±0.52，脂质体包裹组在同期均高于未免疫组和裸DNA免疫组，表明脂质体包裹质粒DNA免疫动物后，能增加动物对病毒感染的抵抗力和反应能力。

HRP was coupled with the fluorochrome FITC and encapsulated with liposome . pEGFP-N1 plasmid (expressing green fluorescent protein) with encapsulation in commercially available liposome was prepared. The eyes were enucleated 1, 4 and 8 weeks after zonule rupture and anterior segments comprising lenses were incubated in medium containing one of these components. Cryo-sections were made and translocation of fluorescent macromolecule from the medium into the lens and green fluorescent protein expression in the epithelium were observed by fluorescent microscopy.

制备FITC标记的HRP脂质体；制备pEGFP-N1质粒（表达绿色荧光蛋白的质粒），并用脂质体包裹；豚鼠悬韧带部分离断后1周、4周、8周取含晶状体的眼前段标本分别在含有FITC-HRP复合物、pEGFP-N1质粒的培养基中孵育，冰冻切片，荧光显微镜观察FITC-HRP复合物进入晶状体和pEGFP-N1质粒在晶状体内表达的情况，比较悬韧带离断侧与非离断侧的差异。3。

Then the combinant plasmid was conducted into ACC-2 cell by electroporation. ACC-2 cells stably expressing CD was obtained by 10-day positive selection with 400 μg/mL G418. Total RNA was extracted and the expression of the CD gene in transfected ACC-2 cells was identified by RT-PCR. RESULTS: PCR yielded a fragment of l280bp and CD was verified by sequence analysis.

测序正确后，将其亚克隆到质粒表达载体pIRES中，构建以内部核糖体进入位点相连的CD基因的质粒表达载体pIRES-CD，采用电穿孔法，以质粒表达载体转染ACC-2细胞，用400μg/mL的G418筛选10d，获得稳定表达CD基因的ACC-2细胞系，提取该细胞的总RNA，用RT-PCR检测CD基因的表达。

The chromatin unfolding assay showed that ,like the wild-type transactivation domain, two variants that represent benign polymorphisms did not induce chromatin unfolding or only induced subtle change. Contrary to the behaviors of the wild type and two benign variants, four cancer-predisposing mutations in the transactivation domain superactivate the chromatin unfolding. The results suggest that the chromatin unfolding assay can aid in the characterization of deleterious mutations in the C-terminal transactivation domain of BRCA1 and may provide more reliable presymptomatic risk assessment.

对这些重组质粒的染色质伸展活性检测表明，野生型pwt和两种良性多态性突变体不具有染色质伸展活性或只有极微弱的染色质伸展活性，而其他4种乳腺癌易感突变体均具有过强的染色质伸展活性，提示利用染色质伸展技术可预测BRCA1转录激活区基因型与乳腺癌发生风险的表现型的关系。

The amplified fragment was inserted into pRSET-A to generate the first double repeat of the precursor gene. By utilizing a pair of isocaudamer BamH I and Bgl II sites, and another downstream Hind III site of plasmid pRSET-A, following a series of simple double digestions and ligation of the resulted products, a series of repeat (3, 4 and 6) precursor peptide fragment genes were derived.

本实验设计一对两侧含编码疏水性氨基酸密码子的引物，经过扩增前导序列10~34aa基因序列，并重新克隆入质粒pRSET-A构建串联二聚体后，再利用质粒pRSET-A的BamH I / Bgl II同尾酶克隆位点，经一系列简单的酶切和连接，快速构建这一前导肽中不含组氨酸标签序列的串联多聚体基因，并成功表达其六聚体重组蛋白。

METHODS: Lipofectamine method: The BMMSCs were obtained from the tibias and femurs of the guinea pigs. The third passage BMMSCs were cultured with plasmid-lipofectamine mixture for 6 hours, followed by fetal bovine medium for 48 hours. Immunohistochemistry was performed for transient expression. G418 was added after 48 hours.

①脂质体法：取体外分离、培养的第3代豚鼠骨髓间充质干细胞，将质粒-脂质体混合物加入含细胞的培养基中培养6 h，再加入胎牛血清的培养基，孵育48 h 后行免疫组织化学检测，即为瞬时表达。48 h后加入含G418培养基筛选。

She gently rebuff ed him, but agreed that they could be friends

她婉言拒绝了，但同意作为朋友相处。

If in the penal farm, you were sure to be criticized.

要是在劳改农场，你等着挨绳子吧！