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A number of CCP domains with the new gene and biological characteristics of


, Military Academy of Medical Sciences, Institute of Microbiology and Epidemiology, pathogenic microorganisms State Key Laboratory Biosafety (Liu Wei, Zhang Fang, Wu Xiaoming, Zhao Qiumin, Zhang Pan River, Yang, Cao Services Spring); Military Medical Sciences, Institute of Biotechnology (Zhu Xudong) Corresponding author: Chun Cao services, E Ⅱ laiI: c spider wc @ nic. . . m,[link widoczny dla zalogowanych], Tel :010-669485345 '-1wIcH' GAC'TC. . 3,[link widoczny dla zalogowanych], reverse primer sequence (P2) is: 5'-GATA. CIEACCTIV_. GTGTGAGA-3, forward and reverse primer containing Iq, nI and Uindm restriction sites. Double digestion (Hind Ⅲ and KpnI) pGI3. Basic vector and PCR product, and then double-digested PCR product was inserted into the vector, that is, the recombinant plasmid 3. Basic. LYP. To build the pGI3. Basic-LYP plasmid-based construct the remaining three haploid recombinant plasmids,[link widoczny dla zalogowanych], using Stratagene's Oui ~ eSite. -DirectedMutagenesis kit, the introduction of H / L, X / Y, P/Q3 positions of single base pair mutation. Primers CHLG1. CHI_G2 (sequences were: 5'-CCAGGCAAGCCTGTGTAAAACACCACA ~ L-qEG-3), the L mutation H, HYP haploid recombinant plasmid obtained, primers CXYG1. CXYG2 (sequences were 5'-CATIT-GTIL-qEACTGCCAC-C-GAAAGCATG plasmid; primers TPQC1. 'rPQC2 (sequences were 5.DGD0 Tong 4rGTTCATTAAC-3 and 5'-IAArGAACA. CATATITACC-AAGCA'IGCCCTCTGTCCYA-3), the mutant P Q, obtained LYQ haploid recombinant plasmid. sequenced that have been successfully constructed plasmids required 4. to build the four kinds of recombinant plasmid (pGI3.basic.HYP, pGI3.basic.LYP, pGI3.basic-LYQ and pGt3.basic.LXP), OGI3.basic and OO3. o0II.trol plasmid with the 0.4gg 16ngpRL-TK plasmid LipofectAMINE20 (D reagent were transfected into NIH3T3 cells. to pGI3.basic as a negative control to pGI3.control as a positive control, using pRL-TK plasmid as an internal reference, each test, each plasmid 3 parallel holes, the same transfection experiment repeated at least 3 times. 48h after transfection, cells were recovered by Pn Midship ega's dual luciferase reporter gene assay kit gene expression report level. analysis by firefly luciferase and Renilla luciferase to determine the ratio between the reporter gene expression. to pGI3.basic as a negative control, the test compared to the ratio of plasmids and repeated tests after 3 obtained mean comparison between groups showed that, pGI3.basic.HYP cells transfected cells transfected luciferase activity was the highest negative control vector (9.40 ± 1.2 times,[link widoczny dla zalogowanych], pGI3.basic-LYP cells followed by luciferase activity, as the negative control vector (8.80 ± 0.76) times, pGI3 a basic.LYQ and 3.basic-LXP of promoter activity lower in the control vector of (5. 05 ± 1.22) and (3.69 ± 1.03) times. 4 promoter activity between haplotypes have some differences, but analysis of variance no significant difference,[link widoczny dla zalogowanych], F = 3.67, P = 0.069. The results that the promoter region of MBL gene mutation can affect promoter activity, MBL gene may be related to the role in disease susceptibility. On the other hand, the difference was not statistically significant, indicating start activity was not able to explain all of the association between the disease, suggesting that there may not detect low levels of MBL gene that led affecting diseases. (Received Et period: 2005-.09-16)
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