Methods:A pairs of primer were designed according to echinococcus granulosus gene fragment clone and sequence analysis special primer as:P1 5'-GGAATGGAGAGAAGTTAC-3',P2 5'-GCAACCTCCGGAACTTGC-3'.
3、 The porcine IL-2 and IL-4 genes were amplified from the library by special primer, the PCR product was cloned into the pMD18-T vector and sequenced.
METHODS: The genotypes of 90 Chinese Han in Guangdong and 104 Sherpas in Tibet were analyzed by sequence special primer polymerase chain reaction(SSP-PCR) sequencing the surfactant protein A gene.
方法:应用序列特异性引物-聚合酶链反应(polymerase chain reac-tion-sequence specific primer,SSP-PCR)方法,对90名广东汉族人和104例西藏夏尔巴人SP-A基因进行检测。
Methods The viral genotype was identified by special primer multiple PCR and the HBV DNA level was determined by quantitative PCR on fluorescence assay.
The results were as follows: 1. The application of SRAP in peanut molecular analysisSRAP (Sequence-related Amplified Polymorphism) is a new marker system based on PCR and preferely amplifies ORFs (open reading frames) by special primer design.
The neutralization titer was 1:8.With the special primer,the major antigen coding region sequences of E2 gene about 250 bp was obtained by RT-PCR. The identity of nucleotide sequence and amino acid sequences of the major antigen coding region sequences of E2 gene of the isolated strains was 79.9%~87.9% and 77.7%~86.6%.
The glutathione peroxidase cDNA has been acquired by reverse transcriplate-polymerase chain reaction (RT-PCR). mRNA was isolated from the culture liver cancer cells firstly, then trancriplate if into cDNA with AMV reverse transcriptase. By using this cDNA as PCR template and two synthetic special primers, we get about 600 bp DNA fragment after 35 circles. The result is identical with our expectation.
The special primers Of p53 exon 7 as wed as HPV16 E6 and E7 ORFs (Opening Rending Frame) were used with PCR, PCR-SSCP technique, and 35 specimens of cervical carcinoma were examined. The results were as follows: ① HPV16 E6, E7 DNA was found in 25/35 specimens (71. 4%),which proved again HPV16 Infection an important event in cervical carcinogenesis. However only 11/35 (31.42% ) bad E6 and E7 ORFs simultaneously, 3/35 (8. 57%) and 11/35 (31. 42% ) had only E6 or E7 respectively. ② No mutation and LOH (Loss...
The special primers Of p53 exon 7 as wed as HPV16 E6 and E7 ORFs (Opening Rending Frame) were used with PCR, PCR-SSCP technique, and 35 specimens of cervical carcinoma were examined. The results were as follows: ① HPV16 E6, E7 DNA was found in 25/35 specimens (71. 4%),which proved again HPV16 Infection an important event in cervical carcinogenesis. However only 11/35 (31.42% ) bad E6 and E7 ORFs simultaneously, 3/35 (8. 57%) and 11/35 (31. 42% ) had only E6 or E7 respectively. ② No mutation and LOH (Loss of Heterozygote) of p53 exon 7 were found in allof 35 specimens. Additionally in the present study, we developed a non-isotopic PCR-SSCP method.
iang Dezhao(Department of Hematology, Hunan Medical University)A gene of human stem cell factor was amplified from cultured human bone marrow stromacells by reverse transcription-PCR(RT-PCR)with a pair of special primers and recombined withthe vectors pKK233-2 and PUC18.Using analysis of restriction endonuclease mapping,PCR andDNA sequencing,the cloned SCF gene was defined.The over-expression and purification of re-combinant SCF gene and its products are still in progress.