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Nutrient-Gene Expression

Poly(A)⁺ RNA Encoding Proteins Capable of Transporting L-Methionine and/or DL-2-Hydroxy-4-(Methylthio) Butanoic Acid Are Present in the Intestinal Mucosa of Broilers¹

Department of Animal and Poultry Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061 and ^* Novus International, Incorporated, St. Charles, MO 63304

²To whom correspondence should be addressed. E-mail: webbk{at}vt.edu.

To investigate the presence of poly(A)⁺ RNA that encode proteins capable of transporting L-methionine (L-Met) and/or DL-2-hydroxy-4-(methylthio) butanoic acid (HMB), Xenopus oocytes were injected with poly(A)⁺ RNA isolated from broiler intestinal mucosa. Healthy oocytes at stage V or VI were collected from Xenopus laevis and microinjected with water, poly(A)⁺ RNA or size-fractioned poly(A)⁺ RNA. The ability of the injected oocytes to take up either L-Met or HMB was examined by incubating oocytes with [methyl-³H]-L-Met or [5-¹⁴C]-HMB. A greater uptake of L-Met (P < 0.01) and HMB (P < 0.05) by oocytes injected with poly(A)⁺ RNA from the duodenum, jejunum and ileum of the small intestine was observed compared with water-injected oocytes. The greatest (P < 0.05) uptake occurred when poly(A)⁺ RNA from the jejunum or ileum was injected. Injections from four different pools of sucrose gradient–fractionated poly(A)⁺ RNA from all three intestinal segments induced (P < 0.01) L-Met uptake. There were three to four different pools of sucrose gradient–fractionated poly(A)⁺ RNA from the duodenum, jejunum and ileum that induced (P < 0.05) HMB uptake. Uptake of HMB was greater at pH 5.5 than at pH 7.5 and was independent of Na⁺. Uptake of L-Met induced by all four poly(A)⁺ RNA pools decreased dramatically when Na⁺ was removed from the uptake buffer, which indicated that the majority of L-Met uptake was Na⁺-dependent. These results indicate that there are multiple sized poly(A)⁺ RNA that encode proteins capable of mediated transport of L-Met and/or HMB present in broiler intestinal mucosa.

KEY WORDS: • Xenopus • chickens • absorption • DL-2-hydroxy-4-(methylthio) butanoic acid • methionine

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