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Cloning of 3′-end full-length cDNA encoding for MT-I and MT-III in yak and analysis of their sequences

Published online by Cambridge University Press:  29 January 2010

Liu Bin ,
Zhang Li-Ping ,
Wu Jian-Ping ,
Ma Bin-Yun ,
Gao Feng-Qin  and
Yan Lian
Liu Bin
Affiliation:
Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China Academy of Agriculture and Stockbreeding Sciences, Inner Mongolia Autonomous Region, Hohhot 010031, China
Zhang Li-Ping*
Affiliation:
Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
Wu Jian-Ping*
Affiliation:
Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
Ma Bin-Yun
Affiliation:
Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
Gao Feng-Qin
Affiliation:
Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China Grassland Research Institute, Chinese Academy of Agricultural Sciences, Hohhot 010010, China
Yan Lian
Affiliation:
Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
*
*Corresponding authors. E-mail: zhangliping@gsau.edu.cn; wujp@gsau.edu.cn
*Corresponding authors. E-mail: zhangliping@gsau.edu.cn; wujp@gsau.edu.cn
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Abstract

Domestic yak (Bos grunniens) metallothionein-I (MT-I) and metallothionein-III (MT-III) cDNA 3′-end full-length sequences (331 and 378 bp; GenBank accession MT-I no. AY758557, MT-III no. DQ492300) from the total RNA of liver and brain tissues were amplified and cloned by reverse transcription polymerase chain reaction (RT-PCR) and 3′-rapid amplification of cDNA ends (RACE) using the primers YMT-I SP and M13 primers M4, and YMT-III SP and M13 primers M4, respectively, which included the MT-I (183 bp) coding sequence, MT-III (207 bp) cDNA coding sequences, and also the tailing signal AATAAA and Poly(A) at the 3′-ends of the MT-I and MT-III, respectively. The analyses showed that the yak cDNA sequence coding for MT-I protein was composed of 61 amino acids, including 20 cysteines having a conserved tripeptide structure, for example C–X–C, C–C–X–C–C, C–X–X–C and so on. The yak cDNA sequence coding for MT-III protein was composed of 68 amino acids, including 19 cysteines having both MT-III-specific conserved tripeptides and the same conserved tripeptides as MT-I, including T, CPCP, GEGAEA and so on. They were all comparatively conservative in molecular evolution. These structures indicated that MT-III had the same physiological functions regarding heavy metals (detoxification and so on) as MT-I, and also different specific functions from MT-I, including inhibition of neuron growth.

Keywords

yakmetallothioneinsmolecular cloning3′-RACEcDNA
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 6 , Issue 3 , December 2009 , pp. 207 - 214
Copyright
Copyright © China Agricultural University 2009

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References

An, JP, Wang, HJ and Han, PP (2005) The physiological function and studying of metallothionein. Journal of Tianshui Normal University 2: 34–37 (in Chinese with English abstract).Google Scholar
Breathnach, R and Chambon, P (1981) Organization and expression of eucaryotic split genes coding for proteins. Annual Review of Biochemistry 50: 349–383.CrossRefGoogle ScholarPubMed
Hamer, DH (1986) Metallothionein. Annual Review of Biochemistry 55: 913–951.CrossRefGoogle ScholarPubMed
Kägi, JHR and Kojima, Y (1991) Chemistry and biochemistry of metallothionein overview of metallothionein. Methods in Enzymology 205: 613–626.CrossRefGoogle Scholar
Kägi, JHR and Schaffer, A (1988) Biochemistry of metallothionein. Biochemistry 27: 8509–8515.CrossRefGoogle ScholarPubMed
Ma, Bin-Yun, Ren, Hong-Wei and Wu, Jian-Ping (2005) Molecular cloning of cDNA encoding for MT-I/II and analyzing of the protein structure in Yak. China Biotechnology 25: 62–68 (in Chinese).Google Scholar
Margoshes, M and Vallee, BL (1957) A cadmium protein from equine kidney cortex. Journal of American Chemical Society 79: 4813–4814.CrossRefGoogle Scholar
Quaife, C, Findley, SD and Erickson, JC (1994) Induction of a new metallothionein isoform (MT-IV) occurs during differentiation of stratified squamous epithelia. Biochemistry 33: 7250–7259.CrossRefGoogle ScholarPubMed
Robinson, NJ, Tommey, AM and Kuske, C (1993) Plant metallothioneins. Biochemistry 295: 1–10.CrossRefGoogle ScholarPubMed
Roesijadi, G (1992) Metallothioneins in metal regulation and toxicity in aquatic animals. Aquatic Toxicology 22: 81.CrossRefGoogle Scholar
Roesijadi, G (1996) Metallothionein and its role in toxic metal regulation. Comparative Biochemistry and Physiology 113: 117.Google Scholar
Suzuki, JS, Kodama, N, Molotkov, A, Aoki, E and Tohyama, C (1998) Isolation and identification of metallothionein isoforms (MT-1 and MT-2) in the rat testis. Biochemistry 334: 695–701.CrossRefGoogle ScholarPubMed
Uchida, Y and Takio, K (1991) The growth inhibitory factor that is deficient in Alzheimer's disease is a 68 amino-acid metallothionein-like protein. Neuron 7: 337–347.CrossRefGoogle ScholarPubMed
Uchida, Y and Yasuo, I (1995) The N-terminal portion of growth inhibitory factor is sufficient for biological activity. Biological Chemistry 2: 3365–3368.CrossRefGoogle Scholar
Uchida, Y, Gomi, F, Masumizu, T and Miura, Y (2002) Growth inhibitory factor prevents neurite extension and the death of cortical neurons caused by high oxygen exposure through radical scavenging. Biological Chemistry 277: 32353–32359.CrossRefGoogle ScholarPubMed
Vallee, BL (1991) Introduction to metallothionein. Methods in Enzymology 205: 3–7.CrossRefGoogle ScholarPubMed
Vallee, BL (1995) The function of metallothionein. Neurochemistry 27: 23–33.CrossRefGoogle ScholarPubMed
Wang, Wen-Qing, Pan, Xian-Ming and Liu, Jie (1994) Research function in brain metallothionein–GIF. Life Sciences 6: 35–36 (in Chinese).Google Scholar
Zhang, Rong-Chang, Wu, Jian-Ping and Han, Jian-Lin (1994) Yak production in China. Asian Livestock: Yak 615: 115–118.Google Scholar