[Cloning and polymorphism analysis of intron 2 of the GH gene in Takifugu].

Size and nucleotide polymorphisms of intron 2 of the growth hormone (GH) gene was studied in 82 individuals from Takifugu obscurus, Takifugu rubripes and Takifugu niphobles with PAGE, SSCP and cloning. Results showed that there were nine length variants (A~I) in these fishes, ranging from 271351 bp, with a frequency of variation of 24.22%. Sequence analysis of the 9 length variants showed the average percentages of the four bases (ATG and C) were 17.15, 20.77, 37.38 and 24.70, respectively, and the difference between GC content (45.47) and AT content (54.53) was not remarkable. Length variation was mainly due to the variable number of microsatellite TCTG repeats (N=20 approximately 40). Further sequence comparison revealed 4 substitution sites: a (C-->A) transversion at nucleotide 103 and 3 transitions 83(C-->T), 101(A-->G) and 296(G-->A), respectively. Phylogenetic analysis of these length variants placed DD and II into one group first, which was subsequently joined by GG and AA, BB and CC, EE and HH were then clustered together, FF was joined last. Thus, intraspecific variations were much smaller than interspecific variations for the intron 2 of the GH gene.

[Analysis of genetic variations at M857 locus of the a1-Fucosy- trans-ferase (FUT1) ORF in pigs].

Enterotoxigenic Escherichia coli F18 (ETEC F18) is the main pathogen that causes edema disease and post-weaning diarrhea in piglets, and a1-fucosytransferase (FUT1) gene has been identified as a receptor gene encoding the receptor for ETEC F18 bacteria. In this study, the method of PCR-RFLP was used to investigate the among 21 breeds including one wild boar breed and 20 western commercial and Chinese native pig breeds (populations). The results showed that none of the individuals in all 21 breeds possessed the resistant AA genotype, the genetic polymorphisms of the FUT1 locus were only detected in two western pig breeds (Duroc and Yorkshire), Lingao pig and hybrid pig breeds, while the wild boar and all the other Chinese pig breeds only possessed the susceptible GG genotype. The results indicated that Chinese native pig breeds, unlike western pig breeds, lack the genetic background on the resistance to ETEC F18 bacteria. This may be owe to their different origination, as the resistance gene to ETEC F18 might be originated from European wild boar. It was also inferred that edema disease and post-weaning diarrhea caused by ETEC F18 had close relationship with the growth speed of pigs.

[Analysis of polymorphisms in exon 14 in porcine Mx1 gene and identification of new mutation points].

By the PCR-RFLP method, the polymorphisms of exon 14 of Mx1 gene were detected in 7 native and foreign pig breeds. Taken together with the analysis of restriction enzyme Hin6 I digestion, there were 6 genotypes and 3 alleles. And, all individuals of Duroc exhibited the AA genotype but the Sutai pigs exhibited all three kinds of genotypes. The BB genotype was detected only in Meishan pigs and their derivate Sutai pigs. Among all pig breeds, the allele B only appeared in Chinese indigenous pig breeds and developed pig breed in this study, and the allele A was dominant allele in all pig breeds except for Songliao black pig. The results of Chi Square test showed that there were abundant polymorphisms in all pig breeds. The gene frequency of Meishan pig and Songliao black pig showed greatly significant difference to other pig breeds (P<0.01), Sutai pig showed greatly significant difference (P<0.01) to other pig breeds except for Pietrain; and Huai pig showed greatly significant differences (P<0.01) with Pietrain and other indigenous Chinese pig breeds, but no difference with Duroc and Yorkshire (P>0.05). Three new mutation points in three genotypes were identified by sequencing comparison analysis of PCR products, two of which resulted in Thr-->Ala and Glu-->Arg respectively, the last one was a silence mutation, the Glu-->Arg mutation point and this silence mutation point only appeared in the individuals with the BB genotype.