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951.
Huitong Zhou Sajee Kunhareang Hua Gong Qian Fang Jiang Hu Yuzhu Luo Jon G.H. Hickford 《Analytical biochemistry》2011,408(2):340
Here we describe a technique we have named polymerase chain reaction stem–loop conformational polymorphism (PCR–SLCP) analysis for screening DNA sequence variation. This technique uses PCR primers with adapters at the 5′ ends to generate amplicons containing inverted terminal repeat sequences. These enable the formation of specific conformers for single-stranded molecules on denaturation, and sequence variation in the loop region may affect the structure of these and their mobilities during electrophoresis. The technique has the ability to resolve sequence variation in amplicons that cannot be resolved using PCR single-strand conformational polymorphism (PCR–SSCP). PCR–SLCP is simple and sensitive, and the results are highly repeatable. 相似文献
952.
A fine balance between CCNL1 and TIMP1 contributes to the development of breast cancer cells 总被引:1,自引:0,他引:1
Li Peng Ma Yanjiao Wang Ai-guo Gong Pengtao Li Jianhua Yang Ju Ouyang Hongsheng Zhang Xichen 《Biochemical and biophysical research communications》2011,(2):3
Cyclin L1 (CCNL1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP1) are candidate genes involved in several types of cancer. However, the expression of CCNL1 and the relationship between CCNL1 and TIMP1 in breast cancer cells is unknown. Using patients’ breast cancer tissues, the expression of CCNL1 and TIMP1 was measured by cDNA microarray and further confirmed by real-time RT-PCR and western blotting. Overexpression or repression of CCNL1 and TIMP1, individually or together, was performed in breast cancer MDA-MB-231 cells by transient transformation methods to investigate their role in breast cancer cell growth. Simultaneously, mRNA and protein expression levels of CCNL1 and TIMP1 were also measured. CCNL1 and TIMP1 expression was significantly elevated in breast cancer tissues compared with that in peri-breast cancer tissues of patients by cDNA microarray and these results were further confirmed by real-time RT-PCR and western blotting. Interestingly, in vitro experiments showed a stimulatory effect of TIMP1 and an inhibitory effect of CCNL1 on growth of MDA-MB-231 cells. Co-expression or co-repression of these two genes did not affect cell growth. Overexpression of CCNL1 and TIMP1 individually induced overexpression of each other. These data demonstrate that there is a fine balance between CCNL1 and TIMP1, which may contribute to breast cancer development. 相似文献
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954.
955.
In order to increase tocopherol content, genes encoding Arabidopsis homogentisate phytyltransferase (HPT) and γ-tocopherol methyltransferase (γ-TMT) were constitutively over-expressed in lettuce
(Lactuca sativa L. var. logifolia), alone or in combination. Over-expression of hpt could increase total tocopherol content, while over-expression of γ-tmt could shift tocopherol composition in favor of α-tocopherol. Transgenic lettuce lines expressing both hpt and γ-tmt produced significantly higher amount of tocopherol and elevated α-/γ-tocopherol ratio compared with non-transgenic control
and transgenic lines harboring a single gene (hpt or γ-tmt). The best line produced eight times more tocopherol than the non-transgenic control and more than twice than hpt single-gene transgenic line. 相似文献
956.
Wazo Myint Qingguo Gong Jinwoo Ahn Rieko Ishima 《Biochemical and biophysical research communications》2011,405(1):19
Sarcoplasmic reticulum Ca2+ ATPase (SERCA) is essential for muscle function by transporting Ca2+ from the cytosol into the sarcoplasmic reticulum through ATP hydrolysis. In this report, the effects of substitution mutations on the isolated SERCA-nucleotide binding domain (SERCA-N) were studied using NMR. 15N–1H HSQC spectra of substitution mutants at the nucleotide binding site, T441A, R560V, and C561A, showed chemical shift changes, primarily in residues adjacent to the mutation sites, indicating only local effects. Further, the patterns of chemical shift changes upon AMP–PNP binding to these mutants were similar to that of the wild type SERCA-N (WT). In contrast to these nucleotide binding site mutants, a mutant found in patients with Darier’s disease, E412G, showed small but significant chemical shift changes throughout the protein and rapid precipitation. However, the AMP–PNP dissociation constant (∼2.5 mM) was similar to that of WT (∼3.8 mM). These results indicate that the E412G mutant retains its catalytic activity but most likely reduces its stability. Our findings provide molecular insight into previous clinical, physiological, and biochemical observations. 相似文献
957.
958.
Dirong Gong Xiaoyu Jia Baolin Wang Feng Wang Chunyu Zhang Liansheng Jiang 《Inorganica chimica acta》2011,373(1):47-53
A series of 2,6-bis(imino)pyridyl iron(III) complexes of the general formula [2,6-(ArNCMe)2C5H3N]FeCl3 (Ar = -C6H5, 3a; 2-MeC6H4, 3b; 2-EtC6H4, 3c; 2-iPrC6H4, 3d; cyclohexyl, 3e; 4-MeC6H4, 3f; 4-iPrC6H4, 3g; 4-FC6H4, 3h and 4-CF3C6H4, 3i), activated by alkylaluminum, MAO or MMAO, have been investigated in 1,3-butadiene polymerization. Iron(III) complex (3a), with the least steric hindrance around the metal center, gives polymer up to 99% in yield in 4 h (butadiene to iron ratio = 1000), and trans-1,4 selectivity about 94.7% at room temperature in toluene, while those (3b-3d) bearing alkyl substituents at the 2-position of each N-aryl ring exhibit much lower catalytic activity and tunable trans-1,4 selectivity. Introduction of an alkyl group at the 4-position (para-position, 3f and 3g) exerts a slightly beneficial effect on the trans-1,4 selectivity, while electronegative groups at the same position (3h and 3i) affect negatively on the activity. The effects of temperature, types of cocatalyst and Al/Fe molar ratio on the polymerization behavior are investigated. More importantly, a mechanism for forming trans-1,4 structure is also proposed. 相似文献
959.
960.
Dan Wang Jing Mao Bin Zhou Xiao-Fu Liao Shi-Qiang Gong Yan Liu Jing-Tao Zhang 《Biotechnology letters》2011,33(1):191-197