Molecular cloning and characterization of <i>SLA‐DR</i> genes in the 133‐family of the Banna mini‐pig inbred line

Zeng, Renpan; Zeng, Yun‐Yun

doi:10.1111/j.1365-2052.2005.01277.x

Cited by 33 publications

(14 citation statements)

References 3 publications

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“…Pieces of acellular bone extracellular matrix 1:2 cm Â 0:5 cm Â 0:5 cm in size were prepared from inbred Banna minipig bones. 19) Briefly, a block of bone from the upper humerus and tibia cancellous bone and rib was cut into 1:0 Â 1:5 cm portions, immersed in phosphate-buffered saline (PBS) for 72 h, and then in 0.05 M Tris-HCl (pH 7.4) that contained 1 mM acetyl-pepstain, 0.5 mM 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF), 2 mg/ml aprotinin, and 100 mM leupeptin for 48 h. Subsequently, the bone fragments were immersed in 3% Triton X-100 for a further 72 h. After washing in distilled water for 12 h, they were incubated with a DNAse/RNAse solution for 12 h. Finally, the bone matrix was freeze dried in a vacuum for 24 h and sterilized with 0 -CTTCCTTAATGTCACGCACGATTTC Co-60 -rays. The basic parameters of the matrix were porosity of 75.8% and a mean pore size of 253 AE 106 mm.…”

Section: Methodsmentioning

confidence: 99%

Bone Regeneration Using an Acellular Extracellular Matrix and Bone Marrow Mesenchymal Stem Cells Expressing Cbfa1

Dong

Ying

Duan

et al. 2009

Bioscience, Biotechnology, and Biochemistry

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Section: Methodsmentioning

confidence: 99%

Bone Regeneration Using an Acellular Extracellular Matrix and Bone Marrow Mesenchymal Stem Cells Expressing Cbfa1

Dong

Ying

Duan

et al. 2009

Bioscience, Biotechnology, and Biochemistry

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“…As the heterozygotic genes were separated and recombined in the process of inbreeding, BMI has already owned six families and eighteen substrains with different phenotypes and genotypes and the inbreeding coefficient is up to 99.95%. Due to its physiological and anatomical similarities to human and good repetitiveness, BMI is a promising animal model for biological study (including male infertility) (Zeng and Zeng, 2005;Wang et al, 2012;Wei et al, 2013). With the aim of a detailed investigation of the TSSK family in porcine, this study isolated the full-length coding sequences (CDS) of BMI TSSKs, conducted sequence analysis, and presented their expression profiles in different tissues and during different developmental stages.…”

Section: Introductionmentioning

confidence: 99%

Cloning, sequence characterization, and expression patterns of members of the porcine TSSK family

Wang¹,

Huo²,

Wang³

et al. 2015

Genet. Mol. Res.

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ABSTRACT. Testis-specific serine kinases (TSSKs) are a family of serine/ threonine kinases highly expressed in the testes that are responsible for regulating many spermatogenesis-related protein activities. Mutations in this family have a positive relationship with oligospermia and azoospermia in human and mouse. Here, five members of the TSSK family from a Banna mini-pig inbred line (BMI) were cloned, sequenced, and characterized. The (2015) full-length coding sequences of BMI TSSKs varied from 807 (TSSK3) to 1095 bp (TSSK1) and encoded 268 to 364 amino acids with molecular weights in the range 30.11 to 41.34 kDa. Following comparison with TSSK4 genes in other species, BMI TSSK4 was found to contain three alternatively spliced variants, inform1, inform 3, and inform 4. BMI TSSK1 and TSSK2 are co-localized on the Sus scrofa chromosome (SSC) 14, and consist of a single exon; TSSK3, TSSK4, and TSSK6 are on SSC6, SSC7, and SSC2, and consist of two, four, and one exon, respectively. Multiple protein sequence alignment and phylogenetic analysis showed that the regions spanning the S_TKc domains were more conserved between pig and other animals: with TSSK1 and TSSK2 and TSSK3 and TSSK6 displaying the greatest degree of homology across species, and the TSSK4 protein clearly distinct from other members. Multi-tissue RT-PCR showed BMI TSSK1, TSSK3, and TSSK4 were only expressed in the testes and seminal vesicle, TSSK2 was confined to testes only, while TSSK6 was expressed widely in adult tissues but was highest in the testes.

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“…The BMI was established after approximately 30 years of consanguineous inbreeding by a Chinese group. The BMI was developed through more than 20 generations with high inbreeding coefficients [4–6]. BMI is considered as an ideal source for pig to human xenotransplantation to solve the serious shortage of donor organs [7–10].…”

Section: Introductionmentioning

confidence: 99%

Highly Efficient Generation of GGTA1 Biallelic Knockout Inbred Mini-Pigs with TALENs

et al. 2013

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Inbred mini-pigs are ideal organ donors for future human xenotransplantations because of their clear genetic background, high homozygosity, and high inbreeding endurance. In this study, we chose fibroblast cells from a highly inbred pig line called Banna mini-pig inbred line (BMI) as donor nuclei for nuclear transfer, combining with transcription activator-like effector nucleases (TALENs) and successfully generated α-1,3-galactosyltransferase (GGTA1) gene biallelic knockout (KO) pigs. To validate the efficiency of TALEN vectors, in vitro-transcribed TALEN mRNAs were microinjected into one-cell stage parthenogenetically activated porcine embryos. The efficiency of indel mutations at the GGTA1-targeting loci was as high as 73.1% (19/26) among the parthenogenetic blastocysts. TALENs were co-transfected into porcine fetal fibroblasts of BMI with a plasmid containing neomycin gene. The targeting efficiency reached 89.5% (187/209) among the survived cell clones after a 10 d selection. More remarkably 27.8% (58/209) of colonies were biallelic KO. Five fibroblast cell lines with biallelic KO were chosen as nuclear donors for somatic cell nuclear transfer (SCNT). Three miniature piglets with biallelic mutations of the GGTA1 gene were achieved. Gal epitopes on the surface of cells from all the three biallelic KO piglets were completely absent. The fibroblasts from the GGTA1 null piglets were more resistant to lysis by pooled complement-preserved normal human serum than those from wild-type pigs. These results indicate that a combination of TALENs technology with SCNT can generate biallelic KO pigs directly with high efficiency. The GGTA1 null piglets with inbred features created in this study can provide a new organ source for xenotransplantation research.

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Molecular cloning and characterization of SLA‐DR genes in the 133‐family of the Banna mini‐pig inbred line

Cited by 33 publications

References 3 publications

Bone Regeneration Using an Acellular Extracellular Matrix and Bone Marrow Mesenchymal Stem Cells Expressing Cbfa1

Bone Regeneration Using an Acellular Extracellular Matrix and Bone Marrow Mesenchymal Stem Cells Expressing Cbfa1

Cloning, sequence characterization, and expression patterns of members of the porcine TSSK family

Highly Efficient Generation of GGTA1 Biallelic Knockout Inbred Mini-Pigs with TALENs

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