PCR-RFLP analysis of mitochondrial DNA ND5/6 region among 3 subspecies of common carp (Cyprinus carpio L.) and its application to genetic discrimination of subspecies

Mitochondrial DNA ND5/6 region was studied by PCR-RFLP analysis among ten representative strains belonging to three subspecies (Cyprinus carpio carpio, Cyprinus carpio haematopterus and Cyprinus carpio rubrofuscus) of common carp (Cyprinus carpio L.). A total of 2.4 kb fragment was amplified and subjected to restriction endonuclease analysis with nine restriction endonucleases subsequently. The results indicated that each subspecies owned one hyplotype and four restriction enzymes (Dde I, HaeIII, Taq I and Mbo I) produced diagnostic restriction sites which could be used for discriminating the three subspecies and as molecular genetic markers for assistant selective breeding of common carp.

Detection of hepatotoxic Microcystis strains by PCR with intact cells from both culture and environmental samples

Microcystins are small hepatotoxic peptides produced by a number of cyanobacteria. They are synthesized non-ribosomally by multifunctional enzyme complex synthetases encoded by the mcy genes. Primers deduced from mcy genes were designed to discriminate between toxic microcystin-producing strains and non-toxic strains. Thus, PCR-mediated detection of mcy genes could be a simple and efficient means to identify potentially harmful genotypes among cyanobacterial populations in bodies of water. We surveyed the distribution of the mcyB gene in different Microcystis strains isolated from Chinese bodies of water and confirmed that PCR can be reliably used to identify toxic strains. By omitting any DNA purification steps, the modified PCR protocol can greatly simplify the process. Cyanobacterial cells enriched from cultures, field samples, or even sediment samples could be used in the PCR assay. This method proved sensitive enough to detect mcyB genes in samples with less than 2,000 Microcystis cells per ml. Its accuracy, specificity and applicability were confirmed by sequencing selected DNA amplicons, as well as by HPLC, ELISA and mouse bioassay as controls for toxin production of every strain used.

Three-piece-ligation PCR and application in disruption of chlorophyll synthesis genes in Synechocystis sp PCC 6803

Linear DNA, consisting of a drug-resistance marker and long flanking sequences, was synthesized by one-step polymerase chain reaction after a three-piece ligating reaction. Chlorophyll synthesis genes, chlH and chIL in Synechocystis sp. PCC 6803, were replaced by a kanamycin-resistance marker through double recombinations with flanking homology regions. Under LAHG conditions, the chIL but not chlH mutant stopped chlorophyll synthesis, while both synthesized chlorophyll in the light.

Enhanced fiber Bragg grating sensor demodulation technique using cascade wavelength division multiplex couplers

An enhanced technique for interrogating fiber Bragg grating wavelength shift using cascade wavelength division multiplexer (WDM) couplers was proposed and demonstrated. Three WDM couplers which show a linear filter function over the expected wavelength range are employed and cascaded to track Bragg wavelength shifts. Compared with single WDM demodulator. sharper spectral slope is obtained and considerable linear filter range is kept. The static and dynamic strain sensor demodulation experiments demonstrated that the simple passive technique improves the sensitivity approximately two times and keeps 5nm linear demodulation range based on our devices. The cascade WDM coupler demodulation system has high scan rate which can be used to monitor fast vibration.

(1) Barnase、Barstar基因和TA29基因5'区调控顺序的PCR扩增、克隆和测序（2）水稻花粉核蛋白因子与TA29基因5'区调控顺序的相互作用

本文运用PCR技术，分别从烟草和B. amyloliquefaciens的基因组DNA中扩增出了TA29基因5'区调控顺序、Barnase和Barstar基因，克隆后进行序列分析，表明其核苷酸顺序与文献报道的一致，然后进行了初步的融合基因的构建。另外，凝胶滞后实验表明，水稻花粉（含绒毡层组织）细胞核中某种蛋白因子能够与TA29基因5'区调控顺序发生特异的结合。本文还对TA29基因5'区调控顺序及Barnase和Barstar基因的潜在应用价值和存在的问题进行了讨论。