• Asian-Australasian Journal of Animal Sciences
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• 제18권9호
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• pp.1226-1230
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• 2005

The present investigation was undertaken to study the genetic polymorphism of the DRB3 exon 2 in 75 crossbred cattle by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Five genotypes i.e. HaeIII-a, HaeIII-b, HaeIII-e, HaeIII-ab and HaeIII-ae were observed when the 284 bp PCR products were digested with HaeIII restriction enzyme. The corresponding frequencies of these patterns were 0.53, 0.04, 0.01, 0.38 and 0.04, respectively. Digestion with RsaI restriction enzyme resolved 24 different restriction patterns. The frequencies of these patterns ranged from 0.013 (RsaI-f, RsaI-k and RsaI-c/n) to 0.120 (RsaI-n). The results revealed that the crossbred cows belonged to the RsaI patterns namely b, k, l, a/l, d/s, l/n, l/o and m/n, whose corresponding frequencies were 0.027, 0.013, 0.040, 0.027, 0.040, 0.067, 0.027 and 0.067, respectively. Digestion of the 284 bp PCR product of DRB3.2 gene with PstI in the crossbred cattle did not reveal any restriction site. These results suggested the absence of the recognition site in some of the animals. These results also revealed that the crossbred cows studied were in homozygous as well as heterozygous condition. On the basis of the above results it can be concluded that the DRB3.2 gene was found to be highly polymorphic in the crossbred cattle population.

• 한국수산과학회지
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• 제32권1호
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• pp.117-120
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• 1999

해산 녹조류 참흩파래, Monostroma nitidum의 엽체를 효소처리하여 다량의 원형질체를 분리하였다. 최적 효소액의 조합은 $4\%$ R-10+$3\%$ Macerozyme R-10+$3\%$ Abalone acetone power로서 생체조직 300mg 당 $4.41\times10^6$개의 원형질체가 분리되었다. 원형질체의 수율은 효소처리 270분에 최대였다. 분리 직후의 원형질체는 구형으로 직경 $13\~33\mu$m의 크기였다. 분리된 원형질체는 0.4M mannitol을 함유한 f/2배지에서 배양한 후 매주 mannitol이 함유되지 않은 f/2 배지로 절반씩 교환함으로서 분화율을 높일 수 있었다. f/2배지를 사용한 적정 배양조건에서 원형질체는 배양 3일 후 새로운 세포벽을 형성하였으며 10일 후 발아하기 시작하여 엽체로 발달하였다. 항생물질의 배지내 첨가는 배양체의 분화를 저해하였다.

• Plant Resources
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• 제7권3호
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• pp.187-194
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• 2004

Sequence-tagged site (STS) markers tightly linked to the bacterial leaf blight (BLB) resistance genes, xa5, xa13 and Xa21, were used in this study. A survey was conducted to find polymorphisms between the resistant and susceptible germplasm in rice. 500 of Korean varieties and 100 of landraces were evaluated in this study. STS marker, RG207 was used to having xa5 resistance gene of rice germplasm. 27 varieties of Korean germplasm showed resistant for xa5 gene. The RG136 an xa-13 marker resulted in a single band of approximately 1kb in all the rice accessions studied. In order to detect polymorphism, digestion of the polymerase chain reaction (PCR) product was performed using a restriction enzyme Hinf Ⅰ. The resistant lines resulted in two bands 0.5kb on digestion with Hinf Ⅰ, while the same enzyme did not digest the PCR product of susceptible lines. No polymorphism was detected in Korean varieties and landraces, indicating that they probably do not contain xa13 gene. pTA248 an Xa-21 marker detected a band of 1kb in the resistant lines and bands of either 750bp or 700bp in the susceptible lines. Among germplasm tested, there are no varieties and landraces with Xa21 resistant gene. The results of the germplasm survey will be useful for the selection of parents in breeding programs aimed at transferring these bacterial blight resistance genes from one varietal background to another.

• Asian-Australasian Journal of Animal Sciences
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• 제16권10호
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• pp.1482-1486
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• 2003

In order to compare effects of feeding systems on rumen fermentation characteristics and nutrient digestion, steers were fed either total mixed ration (TMR) or separate concentrate-roughage ration (CR). Total tract digestibility of nutrients was higher in steers receiving TMR. Especially, DM, ADF and NDF in TMR were digested to a greater extent than those in CR. Rumen pH was not influenced by the feeding systems. Holstein steers on TMR had higher ruminal $NH_3$-N than those on CR. Feeding system did not alter VFA production but TMR feeding resulted in lower A/P ratio. TMR feeding tended to increase the number of bacteria and protozoa in the rumen fluid. Also steers fed TMR generally had higher fiber degrading enzyme activities, which might be the result of increased number of cellulolytic microbes in the rumen of animals on TMR. Our results indicate that TMR may provide more favorable condition for nutrient digestion both in the rumen and in the total tract of steers.

• Asian-Australasian Journal of Animal Sciences
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• 제24권6호
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• pp.752-756
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• 2011

Polymorphism of the second exon of the caprine leukocyte antigen-DRB3 gene (CLA-$DRB3^*02$) was investigated in this study. The 285 bp PCR product of 258 individuals from 10 domestic goat breeds in Southwest China was digested with restriction endonucleases PstI and HaeIII and then genotyped. Three alleles and 4 restriction digestion profiles were distinguished by digestion of the PCR fragment by PstI, and 8 alleles and 13 genotypes by HaeIII. For HaeIII restriction enzyme sites, the Chi-square ($X^2$) test showed that all goat breeds in this study did not fit with the Hardy-Weinberg equilibrium (p<0.01 or p<0.05). The highly polymorphic nature of CLA-$DRB3^*02$ was demonstrated and the ranges of gene heterozygosity (He) and polymorphism information content (PIC) were 0.36-0.63 and 0.32-0.55, respectively. Clustering analysis showed that the 10 goat breeds clustered into two groups and Dazu Black goat had a close genetic relationship with Chengdu Grey, Jintang Black and Nanjiang Yellow goats.

• Parasites, Hosts and Diseases
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• 제47권3호
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• pp.259-264
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• 2009

Genetic polymorphisms of encoding antigen B2 gene (AgB2) in Echinococcus granulosus were studied using PCR-RFLP and DNA sequencing among 20 Egyptian isolates. Five isolates from different host origins (humans, camels, pigs, and sheep) were collected and used. All examined isolates of each host group gave very similar patterns of PCR-RFLP after restriction enzyme digestion with Alul, with the gene size of approximately 140 bp and 240 bp for sheep and human isolates, and approximately 150 bp and 250 bp for pig and camel isolates. No digestion pattern was obtained after incubation of all studied isolates with EcoRI. These results reveal high intra-group homogeneity. DNA sequence analysis highlighted that human infecting strain showed 100% identity with respect to sheep infecting isolate, 96% and 99% with pig and camel infecting isolates, respectively.

• The Plant Pathology Journal
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• 제25권3호
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• pp.236-240
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• 2009

Tylenchulus semipenetrans, citrus nematode is an important phytopathogenic nematode and responsible for serious damage on citrus. However, little information is available about genetic variability of T. semipenetrans among different populations with variation of conventional diagnostic characteristics. In this study, we compared the morphometric and genetic characteristics among different populations. The mature female of T. semipenetrans collected in this study had thicker cuticle than those in the previous studies. In comparative sequence analysis of T. semipenetrans populations obtained from Jeju in Korea, we observed genetic variations within clones generated from single individuals. To determine whether variability among copies of nuclear ribosomal DNA sequences exists in the genome of T. semipenetrans, PCR-RFLP technique from individuals of Korean isolates with MseI and MspI restriction enzymes was used to prove experimentally that all populations have intra-specific variations. Restriction enzyme digestion created several fragments on 3.0% agarose gel corresponding to several haplotypes in all populations, though some populations displayed fragment deletion. The total length of fragments was larger than before digestion, indicating sequence heterogeneity within the genome of T. semipenetrans.

• Asian-Australasian Journal of Animal Sciences
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• 제30권2호
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• pp.262-266
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• 2017

Objective: This study was conducted to screen scrotal hernia in domesticated swine from selected breeders in the Philippines. This defect is associated with a cytosine to thymine mutation in the BCL-2 associated X protein (BAX) gene of swine. Methods: Genetic screening was done by DNA extraction followed by amplification and digestion using polymerase chain reaction-restriction fragment length polymorphism, amplifying the 416 bp region of the BAX gene that was subjected to digestion using the Ear I enzyme. Sequencing was also conducted to validate the results. Results: Results revealed that out of 538 samples tested, 411 (76.4%) of the samples were found to be normal whereas the remaining were carriers of the mutation in which 80 (14.9%) were heterozygous mutants and 47 (8.7%) were homozygous mutants. Pietrain breed was found to have the highest incidence. Conclusion: Having a scrotal hernia eliminates the chances of using the boar as a breeder stock because the following generations arising from it would most likely exhibit herniation. It is therefore advised to establish a genetic screening method for Scrotal Hernia in the Philippines to eliminate the negative gene from the herd.

• 한국식품과학회지
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• 제22권4호
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• pp.466-472
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• 1990

우육에 7.5%의 당을 첨가하고 $121^{\circ}C$에서 가열처리하여 단백질 추출량의 변화를 알아보았다. 갈변반응은 가열시간이 길어짐에 따라 심하게 발생하였으며 glucose 첨가에서 가상 심하게 발생하였다. 단백질의 추출량은 전체적으로 가열시간이 길어질수록 감소하였으며 갈변반응이 심하게 발생한 glucose 첨가구에서 크게 감소하였다. 시료에 peptidase를 작용시켰을 때 단백질 추출량은 거의 변화가 없었으며, chymotrypsin을 작용시켰을때 단백질 추출량이 많이 증가하였으나, glucose 첨가구에서는 약간의 증가만 나타내었고 trypsin을 작용시켰을 때 전체적으로 chymotrysin을 작용시켰을 때보다 단백질 추출량이 많이 증가하는 경향을 나타내었다. Peptidase, chymotrypsin 및 trypsin을 동시에 작용시켰을 때에는 단백질 추출량이 상당히 증가하였지만 갈변반응이 심하게 발생하였던 glucose 첨가구에서는 약간의 증가만 나타내었다.

• The Plant Pathology Journal
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• 제17권4호
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• pp.189-193
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• 2001

The relationships between the phytoplasmas infecting Zizyphus jujuba and Paulownia coreana were investigated by PCR-RELP. The 16S rRNA genes of the phytoplasmas were analyzed and compared with each other after PCR amplification. The amplified bands 1.4 kb in size were analyzed by both restriction digestion and sequencing after cloning into a plasmid vector. In some cases, two different kinds of inserts were observed in the isolates that originated from a single plant. However, many of them appeared to be the amplification products of chloroplastic 16S rRNA gene of host plants. The phytoplasma gene could be differentiated from the chloroplastic gene by restriction digestion of the plasmids carrying the amplification products. Only the recombinant plasmids carrying phytoplasma 16S rRNA gene produced a 1.4 kb band when digested with the enzyme BanII. Of the 52 recombinant plasmids analyzed, 42 appeared to contain inserts that originated from the chloroplastic 16S rRNA gene of the host plants. No variation was detected among 16S rRNA gene of nine phytoplasma isolates infecting Z. jujuba. However, the phytoplasmas infecting Z. jujuba were different from that infecting P. coreana.