Cloning,localization and expression analysis of two fw2.2-like genes in small-and large-fruited pear species

Fruit size is one of the most important agronomic characters,which is mainly determined by cell number and cell size.However,our current knowledge about pear is largely unknown.Through counting of pear mesocarp cells at different stages,we found that the cell number,rather than the cell size,is responsible for the differences between small- and large-fruited cultivars.Fruit weight-2.2（fw2.2） is an important quantitative trait locus（QTL） affecting fruit weight in tomato and functions as a negative regulator in carpel cell division.To get more insights into this QTL in pear fruit development,we isolated two putative homologous fw2.2 genes,which were designated as fw2.2-like（PbFWL） genes.PbFWLs encode Cys-rich proteins with the CCXXXXCPC motif and belong to the PLAC8 superfamily.In addition,results from the subcellular localization indicated that PbFWLs were localized in the plasma membrane.The expression profile of the PbFWL genes by qRT-PCR showed they expressed higher in small-sized fruit cultivar than that in large-sized fruit cultivar during the cell division period.In summary,our data suggest that these two PbFWLs might be negatively related to the cell division in pear fruit.

Chromosomal Localization of a Quail Homeobox Gene: Quox-1

Quox 1 is an Antp like homeobox gene of quail embryo whose expression occurs throughout the developing central nevous system. Using a Dig labeled probe, we localized quox 1 gene on the terminal region of the long arm of quail's chromosome 1 by ISH.

Molecular Markers for Leaf Rust Resistance Gene Lr45 in Wheat Based on AFLP Analysis

Amplified fragment length polymorphism （AFLP） analysis was carried out in Thatcher, near isogenic lines （NILs） canting different genes conferring resistance against wheat leaf rust, and TcLr45 × Thatcher F2 progenies were used to develop markers for Lr45 gene. Sixty AFLP primer combinations were screened and most of them provided clear amplification products, 31 primer combinations displayed polymorphism of TcLr45 in 23 NILs. Two AFLP markers closely linked to the gene Lr45 were acquired： P-AGG/M-GAG261bp, which was found closely linked to the Lr45 locus at a distance of 0.6 cM on one side, and P-ACA/M-GGT105bp, which was found at a distance of 1.3 cM on the other side. The specific hands were cloned and subsequently sequenced. The 261-bp fragment produced by P-AGG/M-GAG showed 86% similarity with the sequence of Vulgate Hort I gene; the 105-bp fragment produced by P-ACA/M-GGT showed 96% similarity with the phosphatidylserine decarboxylase gene of the Triticum monococcum. Both included an open reading frame （ORF）.

Local Delivery of C-myc Antisense Oligodeoxynucleotide by Gelatin Coated Platinium-Iridium Stent to Prevent Restenosis in a Normal Rabbit Carotid Artery

Objectives To investigate the feasibility and effect of local deliveryof c-myc antisense oligodeoxynucleotide (ASODN) by gelatin coated Platinium-Iridium stent to prevent restenosis in a normal rabbit carotid artery. Methods Gelatin coated Platinium-Iridium stent were implanted in the right carotid arteries of 32 rabbits under vision. Animals were randomized to the control group and the treated group receiving c-myc ASODN (n=16 respectively).7, 14, 30,90 days following the stenting procedure, morphometry for caculation of neointimal area and mean neointimal thickness were performed.The expression of c-myc protein was detected by immunohistochemical methods. Results 32 stents were successfully implanted into the right carotid arteries in 32 animals.Morphometric analysis showed that neointimal area and mean neointimal thickness siginificantly increased continuously up to 12 weeks after stent implantation,and at each time point , neointimal area and mean neointimal thickness were siginificantly smaller in the treated group than control group. (P<0.001,respectively).c-myc protein expression was weak positive or negative in treated group and positive in control group. Conclusions Gelatin coated Platinium-Iridium stent mediated local delivery of c- myc ASODN is feasibility , and it can inhibit neointimal hyperplasia to prevent restenosis in a normal rabbit carotid artery.

Distribution of C - myc Antisense Oligonucleotides in Rabbits after Local Delivery by Implanted Gelatin Coated Platinium - Iridium Stent

Objectives To assess thefeasibility, efficiency and tissue distribution of local delivered c - myc antisense oligonucleotides (ASODN) by implanted gelatin coated Platinium - Iridium (Pt -Ir) stent. Methods Gelatin coated Pt - Ir stent which absorbed carboxyfluorescein - 5 - succimidyl ester (FAM) labeled c - myc ASODN were implanted in the right carotid arteries of 6 rabbits under vision. Blood samples were collected at the indicated times. The target artery, left carotid artery, heart , liver and kidney obtained at 45 minutes , 2 hours and 6 hours. The concentration of c - myc ASODN in plasma and tissues were determined by Thin Layer Fluorome-try. Tissue distribution of c - myc ASODN were assessed by fluorescence microscopy. Results At 45 min, 2 h, 6 h, the concentration of FAM labeled c -myc ASODN in target artery was 244. 39, 194. 44, 126. 94(μg/g tissues) respectively, and the delivery efficiency were 44. 4% , 35. 4% and 23. 1% respectively. At the same indicated time point, the plasma concentration was 8. 41, 5. 83, 14. 75 (μg/ml) respectively. Therefore c - myc ASODN concentrations in the target vessel were 29, 33 and 9 -fold higher than that in the plasma. There was circumferential distribution of labeled c - myc in the area of highest fluorescein coinciding with the site of medial dissecting from stent-ing, and the label was most intense in target vessel media harvested at 45 min time point and then dispersed to adventitia. Conclusions Gelatin coated Pt - Ir stent mediated local delivery of c - myc ASODN is feasible and efficient. The localization of ASODN is mainly in target vessel wall.