Short Communication: Genetic linkage map of Cucurbita maxima with molecular and morphological markers.

Cucurbita maxima is one of the most widely cultivated vegetables in China and exhibits distinct morphological characteristics. In this study, genetic linkage analysis with 57 simple-sequence repeats, 21 amplified fragment length polymorphisms, 3 random-amplified polymorphic DNA, and one morphological marker revealed 20 genetic linkage groups of C. maxima covering a genetic distance of 991.5 cM with an average of 12.1 cM between adjacent markers. Genetic linkage analysis identified the simple-sequence repeat marker 'PU078072' 5.9 cM away from the locus 'Rc', which controls rind color. The genetic map in the present study will be useful for better mapping, tagging, and cloning of quantitative trait loci/gene(s) affecting economically important traits and for breeding new varieties of C. maxima through marker-assisted selection.

Myocardial ischemic preconditioning upregulated protein 1(Mipu1):zinc finger protein 667 - a multifunctional KRAB/C2H2 zinc finger protein.

Myocardial ischemic preconditioning upregulated protein 1 (Mipu1) is a newly discovered upregulated gene produced in rats during the myocardial ischemic preconditioning process. Mipu1 cDNA contains a 1824-base pair open reading frame and encodes a 608 amino acid protein with an N-terminal Krüppel-associated box (KRAB) domain and classical zinc finger C2H2 motifs in the C-terminus. Mipu1 protein is located in the cell nucleus. Recent studies found that Mipu1 has a protective effect on the ischemia-reperfusion injury of heart, brain, and other organs. As a nuclear factor, Mipu1 may perform its protective function through directly transcribing and repressing the expression of proapoptotic genes to repress cell apoptosis. In addition, Mipu1 also plays an important role in regulating the gene expression of downstream inflammatory mediators by inhibiting the activation of activator protein-1 and serum response element.

Sequence variations in the FAD2 gene in seeded pumpkins.

Seeded pumpkins are important economic crops; the seeds contain various unsaturated fatty acids, such as oleic acid and linoleic acid, which are crucial for human and animal nutrition. The fatty acid desaturase-2 (FAD2) gene encodes delta-12 desaturase, which converts oleic acid to linoleic acid. However, little is known about sequence variations in FAD2 in seeded pumpkins. Twenty-seven FAD2 clones from 27 accessions of Cucurbita moschata, Cucurbita maxima, Cucurbita pepo, and Cucurbita ficifolia were obtained (totally 1152 bp; a single gene without introns). More than 90% nucleotide identities were detected among the 27 FAD2 clones. Nucleotide substitution, rather than nucleotide insertion and deletion, led to sequence polymorphism in the 27 FAD2 clones. Furthermore, the 27 FAD2 selected clones all encoded the FAD2 enzyme (delta-12 desaturase) with amino acid sequence identities from 91.7 to 100% for 384 amino acids. The same main-function domain between 47 and 329 amino acids was identified. The four species clustered separately based on differences in the sequences that were identified using the unweighted pair group method with arithmetic mean. Geographic origin and species were found to be closely related to sequence variation in FAD2.

Analysis of serum proteome profiles in nasopharyngeal carcinoma with different radiosensitivity.

PURPOSE: The research is endeavored to establish a serum protein fingerprint model for predicting radiosensitivity of nasopharyngeal carcinoma through the analysis of the serum expressed proteins of the pre- and post-treatment nasopharyngeal carcinoma patients treated with radiotherapy. METHODS: Surface-enhanced laser desorption ionization-time of flight-mass spectrometry and CM-10 protein chip were used to detect the serum proteomic patterns of 50 nasopharyngeal carcinoma patients with different radiosensitivity. Thirty-eight of the 50 patients after the treatment were also studied. Biomarker Wizard 3.01 and Biomarker Pattern 5.01 were used in combination to analyze the data and to develop predicting models. RESULTS: At the molecular weight range of 2,000-20,000, the software identified an average of 83 mass peaks between radiation sensitive group and radiation resistant group. 11 protein peaks were significantly different. Of the 83 mass peaks, 4 mass peaks (mass/charge ratio [m/z] 2,575, 3,942, 6,117 and 6,778) were chosen automatically to construct a classification tree. The diagnostic accuracy was 78.0 %. M/z 6,117 and 6,778 of the radiation sensitive group after the treatment trended to those of the radiation resistant group. CONCLUSIONS: The technology of surface-enhanced laser desorption ionization-time of flight-mass spectrometry can be used to screen and identify differentially expressed proteins associated with different radiosensitivity in nasopharyngeal cancer. It should be a very useful tool for predicting radiosensitivity of nasopharyngeal carcinoma.

Genetic association of UBE2B variants with susceptibility to male infertility in a Northeast Chinese population.

The ubiquitin-conjugating enzyme 2B gene (UBE2B) is involved in the regular and symmetric organization of the fibrous sheath of sperm flagella. This study aimed to examine the relationship between single nucleotide polymorphisms (SNPs) in UBE2B and infertility in Northeast Chinese men. We carried out a polymerase chain reaction-restriction fragment length polymorphism analysis for SNPs in 312 fertile males and 388 infertile males in Northeast China. Taking advantage of the high degree of linkage disequilibrium among SNPs surrounding UBE2B (r(2) > 0.90), we selected 2 haplotype-tagging SNPs with a minor allele frequency of 5% or greater (rs17167484: g.-293T>G and rs3777373: g.20016A>G) that captured the majority of the genetic variations in a 40-kbp region of this gene. No significant differences between cases and controls were found in the allelic and genotype distribution of the 2 SNPs. However, the haplotype analysis for the 2 SNPs showed that the GA haplotype was significantly associated with a greater than 3-fold decreased risk of male infertility (P = 0.003). Because the frequency of the GA haplotype (1.1%) is relatively low in Chinese men, such a significant finding may occur by chance, but the results are still significant after multiple comparison adjustments (P = 0.012 after Bonferroni's correction). We conclude that the UBE2B polymorphisms g.-293T>G, g.20016A>G and g.9157A>G are not associated with male infertility, and the GA haplotype is likely a protective factor for male fertility in Northeast Chinese men.

Rapid determination of transgene copy number in tobacco by competitive PCR using a pair of SSR primers.

We developed a straightforward, rapid, and inexpensive method to determine transgene copy number in tobacco. The plasmid (pSSRCopy) used for tobacco transformation contains a simple sequence repeat (SSR) locus, PT1199, which was partially deleted in the middle, a homogenous SSR locus in tobacco K326. A 168-bp segment of the cloned PT1199 was shortened to 95 bp by deleting a 73-bp internal fragment. Using a pair of SSR primers, competitive PCR was amplified from genomic DNA from transgenic tobacco harboring pSSRCopy, and the two expected bands were found. The 168-bp band (SSR-168) corresponds to endogenous PT1199 and the 95-bp band (SSR-95) comes from the integrated pSSRCopy. A single copy of a transgene can be easily distinguished from multiple copies by comparing band densities.